LIGHT Inhibitors For Asthma, Lung and Airway Inflammation, Respiratory, Interstitial, Pulmonary and Fibrotic Disease Treatment

ABSTRACT

Methods of treating inflammatory conditions, disease and disorders are provided. Method include, for example, contacting or administering a sufficient amount of a LIGHT inhibitor to a subject to treat the inflammatory condition, disease or disorder.

RELATED APPLICATIONS

This application claims priority to application Ser. No. 60/973,383,filed Sep. 18, 2007, which application is incorporated by referenceherein in its entirety.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

The invention was supported in part by National Institute of Health RO1Grant AI070535. The government may have certain rights in the invention.

INTRODUCTION

Allergic asthma afflicts 10 million people in the US and is responsiblefor approximately five thousand deaths annually. The prevalence ofasthma has significantly increased over the past few decades along withother allergic diseases, which are characterized by helper T cell 2(Th2) responses. Current therapy primarily includes corticosteroids,bronchodilators, and leukotriene antagonists. A subset of those withmore severe disease have a progressive decline in lung functionattributed to airway remodeling which includes bronchial epithelialmucus metaplasia, airway smooth muscle hypertrophy/hyperplasia,subepithelial fibrosis, and increased angiogenesis. Current asthmatreatment has little impact, if any, on airway remodeling.

Inflammatory cells including T cells, eosinophils, macrophages, and mastcells, as well as structural cells including epithelial, smooth muscle,and fibroblasts have roles in the establishment and maintenance ofremodeling. Growth factors and cytokines that regulate remodelinginclude TGF-

, VEGF, IL-5, IL-9, IL-13, and eotaxin. An understanding of novelmechanisms may lead to much needed therapies that target airwayremodeling and Th2 driven lung inflammation.

LIGHT (TNFSF14, p30 polypeptide) is a protein expressed on activatedCD4/CD8 T cells, dendritic cells (DCs), monocytes, and natural killercells (NK). The binding of LIGHT to herpes virus entry mediator (HVEM),which is expressed on resting T cells, DCs, and monocytes, or thelymphotoxin beta receptor (LTβR), which is expressed on DCs and stromalcells, promotes T cell activation, proliferation, and cytokineproduction. Studies have determined that LIGHT deficient animals have nosignificant abnormalities in the development of lymphoid organs andlymphocytes.

SUMMARY

The invention is based at least in part on the finding that LIGHT (P30polypeptide), a TNF superfamily protein expressed on activated T cells,and other immune cells such as dendritic cells, controls the developmentof airway remodeling and TH2 driven lung responses. Inhibiting orblocking LIGHT from interacting with its receptors, HVEM or LTβR, can beused as an anti-inflammatory, for example, to inhibit or suppressasthmatic inflammation, as well as treat airway remodeling, among otherinflammatory conditions, diseases and disorders. In addition, inhibitingor blocking LIGHT from interacting with its receptors, HVEM or LTβR, isapplicable towards a wide range of chronic and acute fibroproliferativediseases of the lung and airways, including pulmonary fibrosis and COPD(chronic obstructive pulmonary disease), and other tissue and organsystems.

In accordance with the invention, there are provided, methods ofreducing or inhibiting lung or airway inflammation (chronic or acute).In one embodiment, a method includes contacting or administering asufficient amount of an inhibitor of LIGHT (p30 polypeptide) to asubject in need thereof to reduce or inhibit lung or airway inflammationin the subject.

In accordance with the invention, there are also provided, methods oftreating asthma. In one embodiment, a method includes contacting oradministering a sufficient amount of an inhibitor of LIGHT (p30polypeptide) to a subject in need thereof to treat asthma.

In accordance with the invention, there are further provided, methodsfor treating a respiratory, interstitial, pulmonary disease or disorder,and fibrotic diseases and disorders (chronic or acute). In oneembodiment, a method includes contacting or administering a sufficientamount of an inhibitor of LIGHT (p30 polypeptide) to a subject to treatthe respiratory, interstitial, or pulmonary disease or disorder, or thefibrotic disease or disorder.

LIGHT inhibitors include, for example, molecules that bind to LIGHT andinhibit LIGHT binding or interaction with HVEM. LIGHT inhibitors alsoinclude molecules that bind to LIGHT and inhibit LIGHT binding orinteraction with LTβR. LIGHT inhibitors further include molecules thatbind to HVEM and inhibit LIGHT binding or interaction with HVEM. LIGHTinhibitors additionally include molecules that bind to LTβR and inhibitLIGHT binding or interaction with LTβR. LIGHT inhibitors moreoverinclude prodrugs of the foregoing.

Invention methods include contact or administration, in vitro, ex vivoor in vivo (e.g., to a subject in need of treatment). In variousembodiments, lung or airway inflammation, asthma, or a symptom caused byor associated with respiratory, interstitial, or pulmonary disease ordisorder, or the fibrotic disease or disorder is reduced, decreased,inhibited, delayed, halted, or prevented in the subject, locally, orregionally in an area (region), tissue or organ of the subject. Inparticular aspects, a symptom is reduced, decreased, inhibited, delayed,halted, or prevented in a respiratory, interstitial or pulmonary tissueor organ. In another aspect, a method reduces, decreases, inhibits,delays, halts, or prevents inflammation or constriction of lung, airwaysor respiratory mucosum. In yet another embodiment, contacting oradministration in vivo is in a subject that has previously experiencedan asthmatic episode or airway- or broncho-constriction or is in need ofairway- or broncho-dilation.

In accordance with the invention, there are also provided, methods ofinhibiting, reducing or decreasing progression, severity, frequency,duration or probability of one or more symptoms caused by or associatedwith lung or airway inflammation or asthma. In one embodiment, a methodincludes administering to a subject an amount of a LIGHT inhibitorsufficient to inhibit, reduce or decrease progression, severity,frequency, duration or probability of a symptom associated with lung orairway inflammation or asthma. In various aspects, asthma is caused byan allergen or by exercise.

Symptoms include, for example, lung, airway or respiratory mucosuminflammation or tissue damage or remodeling, shortness of breath(dyspnea), rapid breathing (tachypnea), wheezing, stridor, coughing,decreased or reduced lung capacity, chest-tightness, chest pain,prolonged expiration, increased heart rate (tachycardia), runny nose,airway-constriction, decreased lung capacity, or an acute asthmaticepisode, or infiltration of a lung or pulmonary or lymphatic tissue(draining lymph nodes), lymph nodes or airway with immune cells, such asleukocytes and eosinophils, hyperplasia of mucus secreting epithelium,inflammatory lesion of lung, goblet cell hyperplasia, or increased Th2cytokine production (e.g., an interleukin such as IL-4, IL-5, IL-9,IL-13, IL-16, IL-17 or IL-25).

Respiratory diseases can affect the upper or lower respiratory tract.Non-limiting examples include asthma, allergic asthma, bronchiolitis andpleuritis. Additional non-limiting examples include allergic disorders,such as Extrinsic bronchial asthma; Allergic rhinitis; Onchocercaldermatitis; Atopic dermatitis, Drug reactions; Nodules, eosinophilia,rheumatism, dermatitis, and swelling (NERDS); Eosophageal andgastrointestinal allergies. Further non-limiting examples include AirwayObstruction, Apnea, Asbestosis, Atelectasis, Berylliosis,Bronchiectasis, Bronchiolitis, Bronchiolitis Obliterans, OrganizingPneumonia, Bronchitis, Bronchopulmonary Dysplasia, Common Cold, Cough,Empyema, Pleural Empyema, Pleural Epiglottitis, Hemoptysis,Hypertension, Kartagener Syndrome, Meconium Aspiration, PleuralEffusion, Pleurisy, Pneumonia, Pneumothorax, Respiratory DistressSyndrome, Respiratory Hypersensitivity, Respiratory Tract Infections,Rhinoscleroma, Scimitar Syndrome, Severe Acute Respiratory Syndrome,Silicosis, Tracheal Stenosis and Whooping Cough. Still furthernon-limiting examples of respiratory diseases include influenza.

In another embodiment, a method includes administering an amountsufficient to inhibit, reduce or decrease progression, severity,frequency, probability, duration or prevent one or more adversephysiological or psychological symptoms caused by or associated with achronic or acute condition, disorder or disease caused by or associatedwith undesirable or abnormal lung or airway inflammation, asthma, or arespiratory, interstitial, or pulmonary disease or disorder. Inparticular aspects, a condition, disorder or disease is allergic asthma,an acute asthmatic episode, airway constriction, or lung or airwayinflammation, or a respiratory, interstitial, or pulmonary disease ordisorder.

Invention treatment methods include providing a given subject with anobjective or subjective improvement of the condition, disorder ordisease, a symptom caused by or associated with the condition, disorderor disease, or the probability or susceptibility of a subject to thecondition or a symptom caused by or associated with the condition,disorder or disease. In various embodiments, treatment reduces,decreases, inhibits, delays, eliminates or prevents the probability,susceptibility, severity, frequency, or duration of one or more symptomscaused by or associated with the condition, disorder or disease. In aparticular aspect, a method inhibits, reduces or decreases theprobability, severity, frequency, duration or preventing a subject fromhaving an acute asthmatic episode (e.g., an acute asthmatic episodecaused by an allergen, allergic asthma or exercise). In anotherparticular aspect, a method reduces the probability, severity,frequency, duration or delays, halts, or prevents airway-constriction.In additional aspects, treatment improves or increases airway-dilation.In further aspects, a treatment improves asthma, reduces or inhibitslung or airway inflammation, or reduces or inhibits a symptom caused byor associated with a respiratory, interstitial, or pulmonary disease ordisorder.

Candidate subjects for methods of the invention include mammals, such ashumans. Candidate subjects for methods of the invention also includesubjects that are in need of treatment, e.g., any subject that maybenefit from a treatment. Candidate subjects for methods of theinvention therefore include subjects that have or are at risk of havinga condition, disorder or disease caused by or associated with asthma,lung or airway inflammation, or a respiratory, interstitial, orpulmonary disease or disorder. In particular aspects, a subject has beendiagnosed as having asthma, lung or airway inflammation, or arespiratory, interstitial, or pulmonary disease or disorder, or is atrisk of having asthma, lung or airway inflammation, or a respiratory,interstitial, or pulmonary disease or disorder.

Methods of the invention can be practiced by administration or contactwith any dose amount, frequency, delivery route or timing of a LIGHTinhibitor. In particular embodiments, a subject is administered orcontacted a LIGHT inhibitor one, two, three, four or more times hourly,daily, biweekly, weekly, monthly or annually. In additional embodiments,an amount administered is about 0.00001 mg/kg, to about 10,000 mg/kg,about 0.0001 mg/kg, to about 1000 mg/kg, about 0.001 mg/kg, to about 100mg/kg, about 0.01 mg/kg, to about 10 mg/kg, about 0.1 mg/kg, to about 1mg/kg body weight, one, two, three, four, or more times per hour, day,biweekly, week, month or annually. In further embodiments, the amountadministered is less than about 0.00001 mg/kg, one, two, three, four, ormore times per hour, day, biweekly, week, month or annually. Inparticular aspects, the amount is administered substantiallycontemporaneously with, or within about 1-60 minutes, hours, or days ofthe onset of a symptom caused by or associated with asthma, lung orairway inflammation, or a respiratory, interstitial, or pulmonarydisease or disorder.

Methods of the invention include routes of contact or administration ofLIGHT inhibitor locally, regionally and systemically. In a particularembodiment, a LIGHT inhibitor is administered to achieve delivery tolungs, airways, or a lung, airway, respiratory, interstitial, orpulmonary area (region), tissue or organ.

Methods of the invention can be practiced in conjunction with one ormore other treatment protocols or therapeutic regimens. In a particularembodiment, a method includes contacting or administering a second agentor drug to the subject prior to, with or following contacting oradministering LIGHT inhibitor. In particular aspects, a second agent ordrug includes an anti-inflammatory, anti-asthmatic or anti-allergy drug;a hormone or a steroid; an anti-histamine, anti-leukotriene, anti-IgE,anti-α4 integrin, anti-β2 integrin, anti-CCR3 antagonist, β2 agonist oran anti-selectin.

Invention compositions can be formulated as appropriate for practice ofthe methods. In one embodiment, a composition includes a LIGHTinhibitor, and a pharmaceutically acceptable carrier. In a particularaspect, the carrier is a physiologically acceptable gas, liquid, drypowder or an aerosol. In an additional particular aspect, the carrier isa capable of traversing into a lung or airway area (region), tissue ororgan, an interstitial, or pulmonary area (region), tissue or organ, ora mucosal area (region), tissue or organ, or epithelium thereof. In afurther particular aspect, the carrier is lipophilic or non-lipophilic.

Invention compositions can also be included in articles of manufactureor kits appropriate for practice of the invention methods. In oneembodiment, a LIGHT inhibitor is included in an article of manufacture.In one aspect, an article of manufacture is a container having disposedtherein a LIGHT inhibitor. In particular aspects, a container comprisesa canister having disposed therein contents comprising a LIGHTinhibitor, said contents under pressure. In another aspect, a containercomprises an aerosol generator or a spray generator (e.g., an inhaler,nasal sprayer or nebulizer). Exemplary inhalers include metered dose anddry powder inhalers. In a further aspect, an article of manufacture isfor delivery of LIGHT inhibitor to the lungs or airways, for example, anintubation tube or face mask.

In accordance with the invention, further provided are kits. In oneembodiment, a kit includes a LIGHT inhibitor or prodrug thereof. In aparticular aspect, a kit includes a LIGHT inhibitor or prodrug thereofdisposed in an article of manufacture for delivery of the LIGHTinhibitor or prodrug to lung, airways or a respiratory, interstitial, orpulmonary area (region), tissue or organ, optionally with instructionsfor administering said LIGHT inhibitor to a subject. In a particularaspect, a kit includes a second drug (e.g., an anti-inflammatory,anti-asthmatic or anti-allergy drug; a hormone or a steroid; ananti-histamine, anti-leukotriene, anti-IgE, anti-α4 integrin, anti-β2integrin, anti-CCR3 antagonist, β2 agonist, anti-selectin orglucocorticoid; H1-receptor antagonist; or a xanthine drug). In moreparticular aspects, an anti-leukotriene is a cysteinyl-leukotriene(Cys-LT); a β2 agonist is a β2-adrenoceptor.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows reduced eosinophilic lung inflammation in LIGHT-deficientmice after chronic challenge with antigen via the airways. Percent ofbronchoalveolar lavagae eosinophils at 1 day and 3 days after lastairway challenge is shown in the graph. Data are mean eosinophils from 4mice per group for each time point.

FIGS. 2A-2B show reduced peribronchial fibrosis and smooth muscle massin lungs of LIGHT-deficient mice after chronic challenge with antigenvia the airways: A) lung sections stained with trichrome to measurefibrosis and for alpha-smooth muscle actin; and B) bar graphsrepresentative of the area of fibrosis or smooth muscle quantified usingimage analysis with normalization for bronchial size.

FIGS. 3A-3D show that LIGHT-deficient CD4 cells undergo extensiveapoptosis after encountering antigen in vivo: Left (top and bottom)shows antigen-specific CD4 T cells (OT-II) visualized by staining forThy1.2 and CD4; Right (top and bottom) shows the degree of apoptosisoccurring within these CD4 T cell populations determined by co-stainingfor annexin V and 7-AAD after gating on Thy1.2+ cells. The percentagesof early and late apoptotic cells are indicated.

FIGS. 4A-4B show LIGHT-deficient CD4 T cells are defective in promotingasthmatic lung inflammation: A) lung histology, by H&E; sections fromtwo individual antigen-challenged mice receiving wild-type orLIGHT-deficient Th2 (OT-II) cells; and B) total leukocyte and eosinophilcounts by differential cytospin stain; and measurements of IL-5 andIL-13 expression by ELISA in bronchoalveolar samples. First set of datain each case, animals challenged with PBS. Second set, animalschallenged with OVA.

FIGS. 5A-5B show LIGHT-deficient T cells are impaired in accumulating inthe lung and lung-draining lymph nodes (LN) following acute exposure toinhaled antigen (OVA): A) a bar graph representing the LN results; andB) a graph representing the lung results.

FIGS. 6A-6C show LIGHT-deficient T cells do not survive in vivo afterchronic exposure to repetitive allergen (OVA) challenge via the airways:A-C) percentages (left) and absolute numbers (right) of wild-type orLIGHT-deficient CD4 T cells expressing OVA-specific TCR Vα2Vβ5 found inA) Bronchoalveolar lavagae; B) Lung; and C) lung-draining lymph nodes(LDLN).

FIGS. 7A-7B show Eosinophilic lung inflammation is reduced aftertherapeutic treatment with a lymphotoxin beta receptor (LTBR) fusionprotein given during chronic allergen challenge: A) Total infiltratingcells (left) and eosinophils (right) in bronchoalveolar lavage; and B)total infiltrating cells (left) and eosinophils (right) in lung tissue.

DETAILED DESCRIPTION

The invention provides methods of reducing or inhibiting lung or airwayinflammation (chronic or acute). The invention also provides methods oftreating asthma. The invention further provides methods for treating arespiratory, interstitial, pulmonary disease or disorder, and fibroticdiseases and disorders (chronic or acute). In various embodiments, amethod includes contacting or administering a sufficient amount of aninhibitor of LIGHT (p30 polypeptide) to a subject to reduce or inhibitlung or airway inflammation, to treat asthma or to treat therespiratory, interstitial, or pulmonary disease or disorder, or thefibrotic disease or disorder.

The term “an inhibitor of LIGHT,” means a molecule that directly orindirectly inhibits binding of LIGHT (p30 polypeptide) to HVEM or toLTβR. Inhibitors therefore include molecules that bind to LIGHT as wellas molecules that bind to a LIGHT receptor or target. Since LIGHT (p30polypeptide) can bind to a variety of receptors and targets, such asHVEM and LTβR, LIGHT (p30 polypeptide) inhibitors therefore includemolecules that bind to LIGHT (p30 polypeptide), molecules that bind toHVEM, as well as molecules that bind to LTβR, which can thereby inhibitbinding of LIGHT to HVEM, binding of LIGHT to LTβR, etc., eitherdirectly or indirectly.

A non-limiting representative example of human LIGHT (p30 polypeptide)sequence (SEQ ID NO:1; the amino acid residues of the transmembranedomain are shaded, and the amino acid residues of the extracellulardomain are underlined) target for an inhibitor is as set forth below:

MEESVVRPSVFVVDGQTDIPFTRLGRSHRRQSCSVARVGLGLLLLLMGAGLAVQGWFLLQLHWRLGEMVTRLPDGPAGSWEQLIQERRSHEVNPAAHLTGANSSLTGSGGPLLWETQLGLAFLRGLSYHDGALVVTKAGYYYIYSKVQLGGVGCPLGLASTITHGLYKRTPRYPEELELLVSQQSPCGRATSSSRVWWDSSFLGGVVHLEAGEEVVVRVLDERLVRLRDGTRSYFGAFMV

A non-limiting representative example of human HVEM (herpesvirus entrymediator) sequence target for an inhibitor, also referred to as tumornecrosis factor receptor superfamily, member 14 (TNFRSF14) is as setforth below (SEQ ID NO:2):

MEPPGDWGPPPWRSTPKTDVLRLVLYLTFLGAPCYAPALPSCKEDEYPVGSECCPKCSPGYRVKEACGELTGTVCEPCPPGTYIAHLNGLSKCLQCQMCDPAMGLRASRNCSRTENAVCGCSPGHFCIVQDGDHCAACRAYATSSPGQRVQKGGTESQDTLCQNCPPGTFSPNGTLEECQHQTKCSWLVTKAGAGTSSSHWVWWFLSGSLVIVIVCSTVGLIICVKRRKPRGDVVKVIVSVQRKRQEAEGEATVIEALQAPPDVTTVAVEETIPSFTGRSPNH

A non-limiting representative example of human LTβR sequence target foran inhibitor, is as set forth below (SEQ ID NO:3):

MLLPWATSAPGLAWGPLVLGLFGLLAASQPQAVPPYASENQTCRDQEKEYYEPQHRICCSRCPPGTYVSAKCSRIRDTVCATCAENSYNEHWNYLTICQLCRPCDPVMGLEEIAPCTSKRKTQCRCQPGMFCAAWALECTHCELLSDCPPGTEAELKDEVGKGNNHCVPCKAGHFQNTSSPSARCQPHTRCENQGLVEAAPGTAQSDTTCKNPLEPLPPEMSGTMLMLAVLLPLAFFLLLATVFSCIWKSHPSLCRKLGSLLKRRPQGEGPNPVAGSWEPPKAHPYFPDLVQPLLPISGDVSPVSTGLPAAPVLEAGVPQQQSPLDLTREPQLEPGEQSQVAHGTNGIHVTGGSMTITGNIYIYNGPVLGGPPGPGDLPATPEPPYPIPEEGDPGPPGLSTPHQEDGKAWHLAETEHCGATPSNRGPRNQFITHD

Exemplary LIGHT inhibitors include, for example, small organic compounds(e.g., drugs), polypeptide sequences such as antibodies and antibodysubsequences that bind to LIGHT (p30 polypeptide), HVEM (herpesvirusentry mediator) or LTβR (lymphotoxin beta receptor). Additionalexemplary LIGHT inhibitors include, for example, a LIGHT, HVEM or LTβR(lymphotoxin beta receptor) polypeptide subsequence, variant sequence,chimeric sequence or dominant negative sequence (e.g., soluble forms ofLIGHT, HVEM or LTβR). Further exemplary LIGHT inhibitors include, forexample, chimeric sequences, such as a fusion of a LIGHT, HVEM or LTβRpolypeptide sequence (e.g., soluble forms of LIGHT, HVEM or LTβR) and animmunoglobulin (Ig) sequence.

Exemplary LIGHT antibodies include, for example, antibodies that bind tohuman LIGHT. Non-limiting examples of commercially available antibodiesthat bind to human LIGHT include clone T5-39 (BioLegend, San Diego,Calif.), clone 115520 (R&D Systems, Minneapolis, Minn.), clones A-20 andC-20 (Santa Cruz Biotech, Santa Cruz, Calif.), and clone 4E3 (NovusBiologicals, Inc., Littleton, Colo.).

Antibodies include mammalian, human, humanized, humaneered or primatizedforms of heavy or light chain, V_(H) and V_(L), respectively,immunoglobulin (Ig) molecules. An “antibody” means any monoclonal orpolyclonal immunoglobulin molecule, such as IgM, IgG, IgA, IgE, IgD, andany subclass thereof, which includes intact immunoglobulin molecules,two full length heavy chains linked by disulfide bonds to two fulllength light variable domains, V_(H) and V_(L), individually or in anycombination, as well as subsequences, such as Fab, Fab′, (Fab′)₂, Fv,Fd, scFv and sdFv, unless otherwise expressly stated.

An antibody that binds to LIGHT, HVEM or LTβR antibody means that theantibody has affinity for LIGHT, HVEM or LTβR. “Specific binding” iswhere the binding is selective between the two referenced molecules.Thus, specific binding of an antibody for LIGHT, HVEM or LTβR is thatwhich is selective for an epitope present in LIGHT, HVEM or LTβR.Typically, specific binding can be distinguished from non-specific whenthe dissociation constant (K_(D)) is less than about 1×10⁻⁵ M or lessthan about 1×10⁻⁶ M or 1×10⁻⁷ M. Selective binding can be distinguishedfrom non-selective binding using assays known in the art (e.g.,immunoprecipitation, ELISA, Western blotting) with appropriate controls.

Monoclonal antibodies are made by methods known in the art (Kohler etal., Nature, 256:495 (1975); and Harlow and Lane, Using Antibodies: ALaboratory Manual, Cold Spring Harbor Laboratory, 1999). Briefly,monoclonal antibodies can be obtained by injecting mice with antigen.The polypeptide or peptide used to immunize an animal may be derivedfrom translated DNA or chemically synthesized and conjugated to acarrier protein. Commonly used carriers which are chemically coupled tothe immunizing peptide include, for example, keyhole limpet hemocyanin(KLH), thyroglobulin, bovine serum albumin (BSA), and tetanus toxoid.Antibody production is verified by analyzing a serum sample, removingthe spleen to obtain B lymphocytes, fusing the B lymphocytes withmyeloma cells to produce hybridomas, cloning the hybridomas, selectingpositive clones that produce antibodies to the antigen, and isolatingthe antibodies from hybridoma cultures. Monoclonal antibodies can beisolated and purified from hybridoma cultures by a variety ofestablished techniques which include, for example, affinitychromatography with Protein-A Sepharose, size-exclusion chromatography,and ion-exchange chromatography (see e.g., Coligan et al., CurrentProtocols in Immunology sections 2.7.1-2.7.12 and sections 2.9.1-2.9.3;and Barnes et al., “Methods in Molecular Biology,” 10:79-104, HumanaPress (1992)).

A “human antibody” means that the amino acid sequence of the antibody isfully human, i.e., human heavy and light chain variable and constantregions. The antibody amino acids are coded for in the human DNAantibody sequences or exist in a human antibody. Fully human antibodiescan be made by human antibody transgenic or transchromosomic animals,such as mice, or by isolation from human antibody producing cell lines(e.g., B cells) by recombinant DNA methodology known to the skilledartisan, such as gene cloning by reverse transcriptase polymerase chainreaction (RT-PCR). An antibody that is non-human may be made fully humanby substituting non-human amino acid residues with amino acid residuesthat exist in a human antibody. Amino acid residues present in humanantibodies, CDR region maps and human antibody consensus residues areknown in the art (see, e.g., Kabat, Sequences of Proteins ofImmunological Interest, 4^(th) Ed. US Department of Health and HumanServices. Public Health Service (1987); Chothia and Lesk, J. Mol. Biol.(1987) 186:651; Padlan Mol. Immunol. (1994) 31:169; and Padlan Mol.Immunol. (1991) 28:489). Methods of producing human antibodies are alsodescribed, for example, in WO 02/43478 and WO 02/092812.

The term “humanized,” when used in reference to an antibody, means thatthe antibody sequence has non-human amino acid residues of one or morecomplementarity determining regions (CDRs) that specifically bind to theantigen in an acceptor human immunoglobulin molecule, and one or morehuman amino acid residues in the framework region (FR) that flank theCDRs. Any mouse, rat, guinea pig, goat, non-human primate (e.g., ape,chimpanzee, macaque, orangutan, etc.) or other animal antibody may beused as a CDR donor for producing humanized antibody. Human frameworkregion residues can be replaced with corresponding non-human residues(e.g., from the donor variable region). Residues in the human frameworkregions can therefore be substituted with a corresponding residue fromthe non-human CDR donor antibody. A humanized antibody may includeresidues, which are found neither in the human antibody nor in the donorCDR or framework sequences. The use of antibody components derived fromhumanized monoclonal antibodies reduces problems associated with theimmunogenicity of non-human regions. Methods of producing humanizedantibodies are known in the art (see, for example, U.S. Pat. Nos.5,225,539; 5,530,101, 5,565,332 and 5,585,089; Riechmann et al., (1988)Nature 332:323; EP 239,400; WO91/09967; EP 592,106; EP 519,596; PadlanMolecular Immunol. (1991) 28:489; Studnicka et al., Protein Engineering(1994) 7:805; Singer et al., J. Immunol. (1993) 150:2844; and Roguska etal., Proc. Nat'l. Acad. Sci. USA (1994) 91:969).

The term “humaneered,” when used in reference to an antibody, means thatthe antibody sequence has high affinity for antigen but has a greaternumber of human germline sequences than a humanized antibody. Typicallyhumaneered antibody has at least 90% or more human germline sequences.

As used herein, the terms “peptide,” “polypeptide” and “protein” areused interchangeably and refer to two or more amino acids covalentlylinked by an amide bond or non-amide equivalent. Polypeptides includefull length native polypeptide, and “modified” forms such assubsequences, variant sequences, fusion/chimeric sequences anddominant-negative sequences.

Peptides include L- and D-isomers, and combinations thereof. Peptidescan include modifications typically associated with post-translationalprocessing of proteins, for example, cyclization (e.g., disulfide oramide bond), phosphorylation, glycosylation, carboxylation,ubiquitination, myristylation, or lipidation. Modified peptides can haveone or more amino acid residues substituted with another residue, addedto the sequence or deleted from the sequence. Specific examples includeone or more amino acid substitutions, additions or deletions (e.g., 1-3,3-5, 5-10, 10-20, or more).

Subsequences and fragments refer to polypeptides having one or morefewer amino acids in comparison to a reference (e.g., native)polypeptide sequence. An antibody subsequence that specifically binds toLIGHT, HVEM or LTβR can retain at least a part of its binding or LIGHTinhibitory or antagonist activity.

A variant peptide can have a sequence with 50%, 60%, 70%, 75%, 80%, 85%,90%, 95%, 96%, 97%, 98%, or more identity to a reference sequence.Variant sequences include naturally occurring alterations of sequence,due to intra-species polymorphisms or different species, as well asartificially produced alterations of sequence. Sequence homology betweenspecies is in the range of about 70-80%. An amino acid substitution isone example of a variant.

A “conservative substitution” is the replacement of one amino acid by abiologically, chemically or structurally similar residue. Biologicallysimilar means that the substitution is compatible with an activity orfunction of the unsubstituted sequence. Structurally similar means thatthe amino acids have side chains with similar length, such as alanine,glycine and serine, or having similar size. Chemical similarity meansthat the residues have the same charge or are both hydrophilic orhydrophobic. Particular examples include the substitution of onehydrophobic residue, such as isoleucine, valine, leucine or methioninefor another, or the substitution of one polar residue for another, suchas the substitution of arginine for lysine, glutamic for aspartic acids,or glutamine for asparagine, serine for threonine, and the like.

Peptides synthesized and expressed as fusion proteins have one or moreadditional domains linked thereto, and are also referred to as chimericpolypeptides. The additional domain(s) may confer an additional functionupon the sequence. For example, HVEM-IgG or LTβR-IgG fusion proteins canhave LIGHT inhibitory activity.

The term “fusion,” when used in reference to two or more molecules(e.g., polypeptides) means that the molecules are covalently attached. Aparticular example for attachment of two protein sequences is an amidebond or equivalent. The term “chimeric,” and grammatical variationsthereof, when used in reference to a protein, means that the protein iscomprised of one or more heterologous amino acid residues from one ormore different proteins.

The term “heterologous,” when used in reference to a polypeptide, meansthat the polypeptide is not normally contiguous with the otherpolypeptide in its natural environment. Thus, a chimeric polypeptidemeans that a portion of the polypeptide does not exist fused with theother polypeptide in normal cells. In other words, a chimericpolypeptide is a molecule that does not normally exist in nature, i.e.,such a molecule is produced by the hand of man, e.g., artificiallyproduced through recombinant DNA technology.

As used herein, the term “mimetic” refers to a synthetic chemicalcompound which has substantially the same structural and/or functionalcharacteristics as the reference molecule. The mimetic can be entirelycomposed of synthetic, non-natural amino acid analogues, or can be achimeric molecule including one or more natural peptide amino acids andone or more non-natural amino acid analogs. The mimetic can alsoincorporate any number of natural amino acid conservative substitutionsas long as such substitutions do not destroy activity.

Peptide mimetics can contain any combination of non-natural structuralcomponents, which are typically from three structural groups: a) residuelinkage groups other than the natural amide bond (“peptide bond”)linkages; b) non-natural residues in place of naturally occurring aminoacid residues; or c) residues which induce secondary structural mimicry,i.e., induce or stabilize a secondary structure, e.g., a beta turn,gamma turn, beta sheet, alpha helix conformation, and the like. Forexample, a polypeptide can be characterized as a mimetic when one ormore of the residues are joined by chemical means other than an amidebond. Individual peptidomimetic residues can be joined by amide bonds,non-natural and non-amide chemical bonds other chemical bonds orcoupling means including, for example, glutaraldehyde,N-hydroxysuccinimide esters, bifunctional maleimides,N,N′-dicyclohexylcarbodiimide (DCC) or N,N′-diisopropylcarbodiimide(DIC). Linking groups alternative to the amide bond include, forexample, ketomethylene (e.g., —C(═O)—CH₂— for —C(═O)—NH—),aminomethylene (CH₂—NH), ethylene, olefin (CH═CH), ether (CH₂—O),thioether (CH₂—S), tetrazole (CN₄—), thiazole, retroamide, thioamide, orester (see, e.g., Spatola (1983) in Chemistry and Biochemistry of AminoAcids, Peptides and Proteins, Vol. 7, pp 267-357, “Peptide and BackboneModifications,” Marcel Decker, NY).

Peptides and peptidomimetics can be produced and isolated using avariety of methods known in the art. Full length peptides and fragments(subsequences) can be synthesized using chemical methods known in theart (see, e.g., Caruthers, Nucleic Acids Res. Symp. Ser. (1980) 215;Horn, Nucleic Acids Res. Symp. Ser. (1980) 225; and Banga, A. K.,Therapeutic Peptides and Proteins, Formulation, Processing and DeliverySystems (1995) Technomic Publishing Co., Lancaster, Pa.). Peptidesynthesis can be performed using various solid-phase techniques (see,e.g., Roberge, Science (1995) 269:202; Merrifield, Methods Enzymol.(1997) 289:3). Automated synthesis may be achieved, e.g., using apeptide synthesizer.

Individual synthetic residues and polypeptides incorporating mimeticscan be synthesized using a variety of procedures and methodologies knownin the art (see, e.g., Organic Syntheses Collective Volumes, Gilman, etal. (Eds) John Wiley & Sons, Inc., NY). Peptides and peptide mimeticscan also be synthesized using combinatorial methodologies. Techniquesfor generating peptide and peptidomimetic libraries are known, andinclude, for example, multipin, tea bag, and split-couple-mix techniques(see, for example, al-Obeidi, Mol. Biotechnol. (1998) 9:205; Hruby,Curr. Opin. Chem. Biol. (1997) 1:114; Ostergaard, Mol. Divers. (1997)3:17; and Ostresh, Methods Enzymol. (1996) 267:220). Modified peptidescan be further produced by chemical modification methods (see, e.g.,Belousov, Nucleic Acids Res. (1997) 25:3440; Frenkel, Free Radic. Biol.Med. (1995) 19:373; and Blommers, Biochemistry (1994) 33:7886).

Inhibitors of LIGHT therefore include those that can bind selectively aswell as those that bind non-selectively to a ligand or target (e.g.,LIGHT, HVEM, LTβR, etc.) in solution, in solid phase, in vitro, ex vivoor in vivo. As used herein, the term “selective” when used in referenceto a LIGHT inhibitor, means that the inhibitor binds specifically to thetarget entity (e.g., LIGHT, HVEM, LTβR, etc.) and does not significantlybind to a non-ligand or non-target entity. A non-selective inhibitormeans that the inhibitor is not selective for the entity to which itbinds, i.e., it cross-reacts with other entities.

LIGHT inhibitors include variants and derivatives that retain at least apart or all of an activity of the non-variant or non-derivatizedinhibitor. A particular activity (e.g., antagonist or inhibitoryactivity) of a LIGHT inhibitor may be less than or greater than theactivity of a corresponding non-variant or non-derivatized LIGHTinhibitor. For example, a LIGHT inhibitor variant or derivative may haveless or greater activity than non-variant or non-derivatized LIGHTinhibitor.

Non-limiting examples of activities that can be retained, at least inpart, include inhibitory or antagonist activity, binding affinity (e.g.,K_(d)), avidity and binding selectivity (specificity) ornon-selectivity. The variant or derivatized inhibitor can exhibit anactivity (e.g., binding affinity) that is greater or less than acorresponding non-variant or non-derivatized inhibitor, e.g., greater orless inhibitory activity, binding affinity (e.g., K_(d)), avidity orbinding selectivity (specificity) or non-selectivity. For example, “atleast a part” of an activity of an inhibitor can be when the variant orderivatized agent has less of an inhibitory activity, e.g., 10-25%,25-50%, 50-60%, 60-70%, 70-75%, 75-80%, 80-85%, 85-90%, 90-95%, 95-99%,100%, or any percent or numerical value or range or value within suchranges. An activity of an inhibitor can be when the variant orderivatized agent has more inhibitory activity, e.g., 110-125%,125-150%, 150-175%, 175-200%, 200-250%, 250-300%, 300-400%, 400-500%,500-1000%, 1000-2000%, 2000-5000%, or more, or any percent or numericalvalue or range or value within such ranges. At least a part of bindingaffinity of an inhibitor can be when the variant or derivatizedinhibitor has less affinity, e.g., 1-3-fold, 1-5-fold, 2-5 fold,5-10-fold, 5-15-fold, 10-15-fold, 15-20-fold, 20-25-fold, 25-30-fold,30-50-fold, 50-100 fold, 100-500-fold 500-1000-fold, 1000-5000-fold, orless (e.g., K_(d)), or any numerical value or range of values withinsuch ranges. At least a part of binding affinity of an inhibitor can bewhen the variant or derivatized inhibitor has more affinity, e.g.,1-3-fold, 1-5-fold, 2-5 fold, 5-10-fold, 5-15-fold, 10-15-fold,15-20-fold, 20-25-fold, 25-30-fold, 30-50-fold, 50-100 fold,100-500-fold 500-1000-fold, 1000-5000-fold, or more (e.g., K_(d)), orany numerical value or range of values within such ranges.

LIGHT inhibitors can be identified by assays known in the art. Forexample, the amount of activity can be assessed directly, such asmeasuring the particular activity (e.g., inhibitor activity, bindingaffinity, avidity, selectivity (specificity) or non-selectivity). Forexample, a LIGHT inhibitor can be identified by inhibition of HVEM orLTβR mediated lymphocyte activation or cell proliferation. A LIGHTinhibitor can also be identified by change in cell expression of amarker, such as ICAM expression. LIGHT inhibitors can further beidentified by the ability to inhibit binding of purified LIGHT topurified HVEM or LTβR (or HVEM-IgG or LTβR-IgG fusion proteins), forexample, when immobilized on a substrate (e.g., plastic) by ELISA, orwhen any of the molecules are transfected into cells that can beidentified by labeling with the corresponding binding partner by flowcytometry. More particularly, for ELISA assays, plate bound LIGHT can bepre-incubated with LIGHT specific inhibitory molecules and blockade ofreceptor fusion protein binding measured by detection of the binding ofthe Fc fusion protein or lack of binding. Blockade of cell surfaceassociated LIGHT binding to receptors is assessed by pre-incubation ofLIGHT inhibitory molecules with cell lines expressing LIGHT on thesurface followed by addition of receptor Fc fusion proteins. Assessmentof inhibition is measured by detection of binding of the receptor fusionproteins or lack of binding by flow cytometry. Inhibition of LIGHTsignaling in vitro can be determined by inhibiting LIGHT mediatedchemokine secretion from colonic epithelial cells (HT29).

As used herein, the term “the same,” when used in reference to a LIGHTinhibitor means that the activity is within about 50% more than or lessthan the reference inhibitor. The term “substantially the same” whenused in reference to inhibitor activity means that the activity iswithin about 100-500% (2-5-fold) or any percent value or range ofpercent values within such ranges, more than or less than the referenceinhibitor. The same, when used in reference to binding affinity, meansthat the dissociation constant (K_(d)) is within about 1-5-fold, or anynumerical value or range of values within such a range, of thereferenced agent (e.g., 1-5 fold greater affinity or 1-5 fold lessaffinity than the reference agent).

The term “substantially the same” when used in reference to bindingaffinity, means that the dissociation constant (K_(d)) is within about 5to 100 fold, or any numerical value or range of values within such arange, of the reference inhibitor (5-100 fold greater affinity or 5-100fold less affinity than the reference inhibitor). The term “the same,”when used in reference to association constant (K_(a)) is within about 1to 5 fold, or any numerical value or range of values within such arange, of the reference inhibitor (within 1-5 fold greater or 1-5 foldless than the association constant, K_(a)). The term “substantially thesame” when used in reference to association constant (K_(a)), means thatthe association constant is within about 5 to 100 fold greater or less,or any numerical value or range of values within such a range, than theassociation constant, K_(a), of the reference inhibitor (5-100 foldgreater or 5-100 fold less than the reference inhibitor).

Dissociation (K_(d)) constants can be measured using radiolabeledinhibitors in competitive binding assays with increasing amounts ofunlabelled inhibitor to generate saturation curves. The target, ligandor receptor used in the binding assay (e.g., LIGHT, HVEM, or LTβR, etc.)can be expressed in vitro, on cells or be present in extracts.Association (K_(a)) and dissociation (K_(d)) constants can be measuredusing surface plasmon resonance (SPR) (Rich and Myszka, Curr. Opini.Biotechnol. 11:54 (2000); Englebienne, Analyst. 123:1599 (1998)). SPRmethods for real time detection and monitoring of protein binding ratesare known and are commercially available and can be used to determinedissociation (K_(d)) constants (BiaCore 2000, Biacore AB, Upsala,Sweden; and Malmqvist, Biochem. Soc. Trans. 27:335 (1999)).

As used herein, the term “contact” and grammatical variations thereofmeans a physical or functional interaction between one entity and one ormore other entities. An example of physical contact is a direct orindirect binding, such as between a LIGHT inhibitor and a target orreceptor. An example of a functional interaction is where anintermediate facilitates or mediates a change in activity of one entityby another entity, such as a signaling pathway where molecules withinthe pathway functionally interact but need not physically contact eachother. In the methods, contact can occur in solution, in solid phase, invitro, ex vivo or in vivo (i.e., in a subject).

In accordance with the invention, there are provided methods insolution, in solid phase, in vitro, ex vivo or in vivo (i.e., in asubject). In one embodiment, a method includes contacting oradministering to a subject, e.g. a subject in need thereof, an amount ofa LIGHT inhibitor to treat the subject. In one particular aspect, anamount of LIGHT inhibitor contacted with or administered to the subjectis sufficient to reduce or inhibit lung or airway inflammation. Inanother particular aspect, an amount of LIGHT inhibitor contacted withor administered to the subject is sufficient to treat asthma. In afurther aspect, an amount of LIGHT inhibitor is administered to asubject sufficient to treat a respiratory, interstitial, or pulmonarydisease or disorder, or fibrotic disease or disorder. In a still furtheraspect, an amount of LIGHT inhibitor is administered to a subject whomhas previously experienced an asthmatic episode or airway-constrictionor obstruction, or is in need of airway-dilation, sufficient to inhibitor reduce airway-constriction or obstruction, or to increase, stimulateor improve airway-dilation.

As used herein, the term “associated with,” when used in reference tothe relationship between a symptom and a condition, disorder or disease,means that the symptom is caused by the referenced condition, disorderor disease, or is a secondary effect of the referenced condition,disorder or disease. A symptom that is present in a subject maytherefore be the direct result of or caused by the referenced condition,or may be due at least in part to the subject reacting or responding tothe referenced condition, disorder or disease, e.g., a secondary effect.For example, symptoms that occur during an asthmatic or allergic episodeare due in part to hypersensitivity or an aberrant response of theimmune system of the subject to the antigen/allergen.

As used herein, the term “subject” includes animals, typically mammaliananimals, such as but not limited to humans, non-human primates (apes,gibbons, chimpanzees, orangutans, macaques), domestic animals (dogs andcats), farm animals (horses, cows, goats, sheep, pigs), and experimentalanimals (mouse, rat, rabbit, guinea pig). Subjects include animaldisease models (e.g., asthma, allergy). Subjects include naturallyoccurring or non-naturally occurring mutated or non-human geneticallyengineered (e.g., transgenic or knockout) animals. Subjects furtherinclude animals having or at risk of having a chronic or acutecondition, disorder or disease.

Conditions, disorders and diseases treatable in accordance with theinvention include, for example, chronic or acute inflammatoryconditions, disorders and diseases, allergies, allergic conditions,disorders and diseases. An “inflammatory” condition, disorder or diseaserefers to one or more physiological responses that characterize orconstitute inflammation. An “allergy” or “allergic condition,” as usedherein refers to a hypersensitivity to a substance (e.g., an allergen).Allergic conditions, disorders and diseases include but are not limitedto allergic asthma, hayfever (seasonal rhinitis), allergic rhinitis,allergic conjunctivitis, eczema, urticaria, food allergies, and otheratopic conditions.

Inflammatory, allergic and non-allergic and conditions, disorders anddiseases of the respiratory system, including airways and lung, includeasthma, chronic obstructive pulmonary disease (“COPD”), granulomatousdiseases of the lungs and lower airway passages, non-malignantproliferative disease of the lungs e.g., idiopathic pulmonary fibrosis,hypersensitivity pneumonitis and bronchopulmonary dysplasia.Non-limiting examples of allergic conditions, disorders and diseasesinclude, for example, extrinsic bronchial asthma; allergic rhinitis(AR); Onchocercal dermatitis; atopic dermatitis, drug reactions;nodules, eosinophilia, rheumatism, dermatitis, and swelling (NERDS);esophageal and gastro-intestinal (GI) allergies.

In accordance with the invention, there are provided methods of reducingprogression, severity, frequency, duration, susceptibility orprobability of inflammatory, allergic and non-allergic conditions,disorders and diseases of the respiratory system. In one embodiment, amethod includes administering to a subject an amount of LIGHT inhibitorsufficient to reduce or decrease progression, severity, frequency,duration, susceptibility or probability of one or more adverse symptomsassociated with inflammation in the respiratory tissue or organ.

In another embodiment, a method includes administering to a subject anamount of LIGHT inhibitor sufficient to reduce or decrease progression,severity, frequency, duration, susceptibility or probability of one ormore adverse symptoms caused by or associated with asthma (allergic ornon-allergic). In one aspect, the adverse symptom is selected from lung,airway or respiratory mucosum inflammation or tissue damage, shortnessof breath, wheezing, coughing, chest-tightness, chest pain, increasedheart rate, runny nose, airway-constriction, decreased lung capacity,and an acute asthmatic episode. In another aspect, asthma is caused byor associated with exposure to an allergen or associated with exercise.In yet another aspect, the subject has been diagnosed as having asthma.

“Asthma” refers to an allergic or non-allergic condition, disorder ordisease of the respiratory system that is episodic and characterized byinflammation with constriction, narrowing or obstruction of the airways.Allergic asthma is typically associated with increased reactivity ofrespiratory system (airways, lung, etc.) to an inhaled agent. Asthma isfrequently, although not exclusively associated with atopic or allergicsymptoms. Typically, a subject with asthma suffers from recurrentattacks of paroxysmal dyspnea (i.e., “reversible obstructive airwaypassage disease”), cough, shortness of breath with wheezing due tospasmodic contraction of the bronchi, sometimes referred to as“bronchospasm,” chest pain, chest tightness, etc. While a plurality ofsuch adverse symptoms typically occur in asthma, the existence of anyone is usually adequate for diagnosis of asthma, and for treatment inaccordance with the invention.

Asthmatic conditions include allergic asthma as well as bronchialallergy, which typically are provoked by a variety of factors includingexercise such as vigorous exercise (“exercise-induced bronchospasm”),and irritant particles (allergens such as pollen, dust, venoms, cotton,dander, foods). Asthmatic conditions can be acute, chronic, mild,moderate or severe asthma (unstable asthma), nocturnal asthma or asthmaassociated with psychologic stress.

“Allergic rhinitis” is an allergic reaction of the nasal mucosa (upperairways), which includes hay fever (seasonal allergic rhinitis) andperennial rhinitis (non-seasonal allergic rhinitis) which are typicallycharacterized by seasonal or perennial sneezing, rhinorrhea, nasalcongestion, pruritis and eye itching, redness and tearing. “Non-allergicrhinitis” refers to eosinophilic non-allergic rhinitis, in subjects withnegative skin tests, and subjects who have abnormal or undesirablenumbers of eosinophils in their nasal secretions.

An “allergen” is a substance that can promote, stimulate or induce anallergic or asthmatic episode in a subject. Allergens include plant/treepollens, insect venoms, animal dander, house dust mite, dust, fungalspores, latex, food and drugs (e.g., penicillin). Examples of particularallergens include proteins specific to the following genera: Canis(Canis familiaris); Dermatophagoides (e.g., Dermatophagoides farinae);Felis (Felis domesticus); Ambrosia (Ambrosia artemiisfolia); Lolium(e.g., Lolium perenne or Lolium multiflorum); Cryptomeria (Cryptomeriajaponica); Alternaria (Alternaria alternata); Alder, Alnus (Alnusgultinosa); Betula (Betula verrucosa); Quercus (Quercus alba); Olea(Olea europa); Artemisia (Artemisia vulgaris); Plantago (e.g., Plantagolanceolata); Parietaria (e.g., Parietaria officinalis or Parietariajudaica); Blattella (e.g., Blattella germanica); Apis (e.g., Apismultiflorum); Cupressus (e.g., Cupressus sempervirens, Cupressusarizonica and Cupressus macrocarpa); Juniperus (e.g., Juniperussabinoides, Juniperus virginiana, Juniperus communis and Juniperusashei); Thuya (e.g., Thuya orientalis); Chamaecyparis (e.g.,Chamaecyparis obtusa); Periplaneta (e.g., Periplaneta americana);Agropyron (e.g., Agropyron repens); Secale (e.g., Secale cereale);Triticum (e.g., Triticum aestivum); Dactylis (e.g., Dactylis glomerata);Festuca (e.g., Festuca elatior); Poa (e.g., Poa pratensisor Poacompressa); Avena (e.g., Avena sativa); Holcus (e.g., Holcus lanatus);Anthoxanthum (e.g., Anthoxanthum odoratum); Arrhenatherum (e.g.,Arrhenatherum elatius); Agrostis (e.g., Agrostis alba); Phleum (e.g.,Phleum pratense); Phalaris (e.g., Phalaris arundinacea); Paspalum (e.g.,Paspalum notatum); Sorghum (e.g., Sorghum halepensis); and Bromus (e.g.,Bromus inermis). Allergens also include peptides and polypeptides usedin experimental animal models of allergy and asthma, including ovalbumin(OVA) and Schistosoma mansoni egg antigen.

A “respiratory disorder” or a “respiratory mucosum disorder” means acondition, disorder or disease related to a tissue or organ of therespiratory system. Examples include, but are not limited to, upper orlower airway inflammation, allergy(ies), breathing difficulty, cysticfibrosis (CF), allergic rhinitis (AR), Acute Respiratory DistressSyndrome (ARDS), pulmonary hypertension, lung inflammation, bronchitis,airway obstruction, airway constriction, airway narrowing,broncho-constriction and inflammation associated with microbial or viralinfections, such as influenza, picornaviridae (rhinoviruses such ashuman rhinovirus (HRV); enteroviruses (EV) such as polioviruses,coxsackieviruses and echoviruses) or severe acute respiratory syndrome(SARS). Additional non-limiting examples of respiratory disorders andrespiratory mucosum disorders include apnea, asbestosis, atelectasis,berylliosis, bronchiectasis, bronchiolitis, bronchiolitis obliteransOrganizing Pneumonia, Bronchitis, Bronchopulmonary Dysplasia, CommonCold, Cough, Empyema, Pleural Empyema, Pleural Epiglottitis, Hemoptysis,Hypertension, Kartagener Syndrome, Meconium Aspiration, PleuralEffusion, Pleurisy, Pneumonia, Pneumothorax, Respiratory DistressSyndrome, Respiratory Hypersensitivity, Respiratory Tract Infections,Rhinoscleroma, Scimitar Syndrome, Severe Acute Respiratory Syndrome(SARS), Silicosis, Tracheal Stenosis, and Whooping Cough.

Further non-limiting examples of interstitial and pulmonary disordersinclude Eosinophilic pleural effusions; Transient pulmonary eosinophilicinfiltrates (Löffler); Histiocytosis; Chronic eosinophilic pneumonia;Hypersensitivity pneumonitis; Allergic bronchopulmonary aspergillosis;Sarcoidosis; Idiopathic pulmonary fibrosis; pulmonary edema; pulmonaryembolism; pulmonary emphysema; Pulmonary Hyperventilation; PulmonaryAlveolar Proteinosis; Chronic Obstructive Pulmonary Disease;Interstitial Lung Diseases; and Topical eosinophilia.

The term “airway,” as used herein, means a part of or the wholerespiratory system of a subject that is exposed to air. “Airways”therefore include the upper and lower airway passages, within which arenot limited to the trachea, bronchi, bronchioles, terminal andrespiratory bronchioles, alveolar ducts and alveolar sacs. Airwaysinclude sinuses, nasal passages, nasal mucosum and nasal epithelium. Theairway also includes, but is not limited to throat, larynx,tracheobronchial tree and tonsils.

Particular non-limiting examples of subjects include subjects having orat risk of having inflammation or lung or airways, an inflammatory orallergic condition, disorder or disease. Non-limiting examples ofsubjects further include subjects having or at risk of having an adverseor undesirable symptom associated with an inflammatory or allergiccondition, disorder or disease, such as asthma. Such at risk subjectscan be identified by a personal or family history, through geneticscreening, tests appropriate for detection of increased risk, orexhibiting relevant symptoms indicating predisposition orsusceptibility.

Subjects having or at risk of having an allergic condition, disorder ordisease include subjects with an existing allergic condition or a knownor a suspected predisposition towards developing a symptom associatedwith or caused by an allergic condition. Thus, the subject can have anactive chronic allergic condition, disorder or disease, an acuteallergic episode, or a latent allergic condition, disorder or disease.Certain allergic conditions, are associated with seasonal orgeographical environmental factors. Thus, at risk subjects include thoseat risk from suffering from a condition based upon a prior personal orfamily history, and the season or physical location, but which thecondition or a symptom associated with the condition may not presentlymanifest itself in the subject.

A subject having or at risk of having asthma refers to a subjectsuffering from an acute episode of asthma, either a new-onset or arecurrent episode, a subject with a prior history of one or moreepisodes of asthma, or a subject with a known or suspectedpredisposition towards developing asthma. A subject having asthma canhave active asthma or can be asymptomatic and between acute asthmaepisodes. A subject having asthma can be suffering from recently acuteasthmatic episode (e.g., within minutes or hours of episode onset). Asubject having asthma can have a positive skin test, or exhibit one ormore symptoms typically associated with acute or chronic asthma, forexample, a symptom of allergic asthma. A subject having or at risk ofhaving asthma may be or has been exposed to an allergen, for example,and is at increased risk of suffering from an asthmatic episode due to apredisposition or susceptibility towards an asthmatic episode uponre-exposure to the allergen. Subjects predisposed or susceptible to,exposed to or allergic to these or other allergens are at risk of havingasthma and, therefore, are amenable to treatment in accordance with theinvention.

At risk subjects also appropriate for treatment in accordance with theinvention include subjects exposed to an allergen or are susceptible tohaving an allergic reaction, or infection or exposure by an agent thatis associated with an allergy or allergic reaction. At risk subjectsappropriate for treatment in accordance with the invention includesubjects having a predisposition towards an allergic reaction, orinfection or exposure to an agent that is associated with an allergy orallergic reaction due to a genetic or environmental risk factor. Methodsof the invention include subjects contacted with or administered to abinding agent prophylactically.

In the methods of the invention in which a detectable result orbeneficial effect is a desired outcome, such as a therapeutic benefit ina subject treated in accordance with the invention, compositions such asLIGHT inhibitors can be administered in sufficient or effective amounts.An “amount sufficient” or “amount effective” includes an amount that, ina given subject, can have a desired outcome or effect. The “amountsufficient” or “amount effective” can be an amount of a LIGHT inhibitorthat provides, in single or multiple doses, alone or in combination withone or more other (second) compounds or agents (e.g., a drug),treatments or therapeutic regimens, a long or short term detectableresponse, a desired outcome or beneficial effect in a particular givensubject of any measurable or detectable degree or duration (e.g., forminutes, hours, days, months, years, or cured).

An amount sufficient or an amount effective can but need not be providedin a single administration and can but need not be administered alone(i.e., without a second drug, agent, treatment or therapeutic regimen),or in combination with another compound, agent, treatment or therapeuticregimen. In addition, an amount sufficient or an amount effective neednot be sufficient or effective if given in single or multiple doseswithout a second compound, agent, treatment or therapeutic regimen,since additional doses, amounts or duration above and beyond such doses,or additional drugs, agents, treatment or therapeutic regimens may beincluded in order to be effective or sufficient in a given subject.Further, an amount sufficient or an amount effective need not beeffective in each and every subject, nor a majority of subjects in agiven group or population. Thus, as some subjects may not benefit fromsuch treatments an amount sufficient or an amount effective meanssufficiency or effectiveness in a particular subject, not a group or thegeneral population. As is typical for such methods, some subjects willexhibit a greater or less response to a method of the invention,including treatment/therapy.

Reducing, inhibiting decreasing, eliminating, delaying, halting orpreventing a progression or worsening or an adverse symptom of thecondition, disorder or disease is a satisfactory outcome. The doseamount, frequency or duration may be proportionally increased orreduced, as indicated by the status of the condition, disorder ordisease being treated, or any adverse side effects of the treatment ortherapy. Dose amounts, frequencies or duration also consideredsufficient and effective are those that result in a reduction of the useof another drug, agent, treatment or therapeutic regimen or protocol.For example, a LIGHT inhibitor is considered as having a beneficial ortherapeutic effect if contact, administration or delivery in vivoresults in the use of a lesser amount, frequency or duration of anotherdrug, agent, treatment or therapeutic regimen or protocol to treat thecondition, disorder or disease, or an adverse symptom thereof.

An “amount sufficient” or “amount effective” includes reducing,preventing, delaying or inhibiting onset, reducing, inhibiting,delaying, preventing or halting the progression or worsening of,reducing, relieving, alleviating the severity, frequency, duration,susceptibility or probability of one or more adverse or undesirablesymptoms associated with the condition, disorder or disease of thesubject. In addition, hastening a subject's recovery from one or moreadverse or undesirable symptoms associated with the condition, disorderor disease is considered to be an amount sufficient or effective.Various beneficial effects and indicia of therapeutic benefit are as setforth herein and are known to the skilled artisan.

An “amount sufficient” or “amount effective,” in the appropriatecontext, can refer to therapeutic or prophylactic amounts.Therapeutically or prophylactically sufficient or effective amounts meanan amount that, in a given subject, detectably improves the condition,disorder or disease, such as an inflammatory condition, disorder ordisease, as assessed by one or more objective or subjective clinicalendpoints appropriate for the condition, disorder or disease.

In accordance with the invention, there are provided methods whichprovide a beneficial effect, such as a therapeutic benefit, to asubject. In one embodiment, a method includes administering an amount ofLIGHT inhibitor sufficient to provide a therapeutic benefit orbeneficial effect to a subject. In one aspect, a method reduces orinhibits probability, susceptibility, severity, frequency, duration orprevents lung or airway inflammation in the subject. In another aspect,a method reduces the probability, susceptibility, severity, frequency,duration or prevents an asthmatic episode (e.g., associated withallergic or non-allergic asthma) in the subject. In an additionalaspect, a method reduces or inhibits probability, susceptibility,severity, frequency, duration or prevents a symptom caused by orassociated with a respiratory, interstitial or pulmonary disease ordisorder. In a further aspect, a method increases, stimulates, enhances,induces or promotes airway-dilation in the subject. In still anotheraspect, a method reduces the probability, susceptibility, severity,frequency, duration or prevents or eliminates airway-constriction thesubject. In a yet further aspect, a method is sufficient to reduceprogression, severity, frequency, duration, susceptibility, probability,halt, eliminate or prevent one or more adverse physiological orpsychological symptoms associated with asthma (allergic ornon-allergic).

Sufficiency or effectiveness of a particular treatment can beascertained by various clinical indicia and endpoints. For example, inorder to ascertain an improvement in asthma, an increase in airwaydilation, lung function or a reduction in airway constriction,obstruction or narrowing, progression, severity, duration, frequency,susceptibility or probability of one or more symptoms of asthma. An“amount sufficient” or “amount effective” to treat asthma is thereforean amount that provides an objective or subjective reduction orimprovement in progression, severity, frequency, susceptibility orprobability of lung or airway inflammation, lung or airway tissuedamage, shortness of breath, wheezing, coughing, chest-tightness, chestpain, increased heart rate, runny nose, airway or broncho-constriction,-obstruction or narrowing, decreased lung capacity, acute asthmaticepisodes, nighttime awakenings, etc. Thus, a reduction, decrease,inhibition, delay, halt, prevention or elimination of one or moreadverse symptoms (e.g., shortness of breath, wheezing, coughing,chest-tightness, chest pain, increased heart rate, runny nose, acuteasthmatic episodes, nighttime awakenings, etc.) can be used as a measureof sufficiency or effectiveness.

A method to determine an improvement in lung or pulmonary function is tomeasure the forced expiratory volume in one second (FEV₁) an increase ofwhich indicates an improvement. Spirometry is a test which measures theamount and rate at which air can pass through airways. Airway narrowingdue to inflammation restricts air flow through the airways, which isdetected by changed spirometry values. Exercise challenge andmethacholine inhalation tests are also used to evaluate airway narrowingor constriction. Yet another method to determine an improvement is tomeasure serum IgE in a subject. A reduction in serum or bronchoalveolarlavage (BAL) fluid IgE is an objective measure of treatment efficacy.Various additional methods are known to the skilled artisan fordetecting improvement in lung or pulmonary function.

An “amount sufficient” or “amount effective” also includes an amountthat, when used in combination with another binding agent, drug, ortreatment or therapeutic regimen, reduces the dosage frequency, dosageamount, or an adverse symptom or side effect of the other binding agent,drug or treatment or therapeutic regimen, or eliminates the need for theother binding agent, drug or treatment or therapeutic regimen. Forexample, an “amount sufficient” or “amount effective” of a LIGHTinhibitor could result in a reduction in the dosage frequency or dosageamount of a steroid, antihistamine, beta adrenergic agonist,anticholinergic, methylxanthine, anti-IgE, anti-leukotriene, anti-beta2integrin, anti-CCR3 antagonist, or anti-selectin required to achieve thesame clinical endpoint.

The terms “treat,” “therapy” and grammatical variations thereof whenused in reference to a method means the method provides an objective orsubjective (perceived) improvement in a subjects' condition, disorder ordisease, or an adverse symptom associated with the condition, disorderor disease. Non-limiting examples of an improvement can therefore reduceor decrease the probability, susceptibility or likelihood that thesubject so treated will manifest one or more symptoms of the condition,disorder or disease. Additional symptoms and physiological orpsychological responses caused by or associated with conditions,disorders or diseases associated with, for example, lung and airwayinflammation, asthma and a respiratory, interstitial or pulmonarydisease or disorder are set forth herein and known in the art and,therefore, improvements in these and other adverse symptoms orphysiological or psychological responses can also be included in themethods of the invention.

Methods of the invention therefore include providing a detectable ormeasurable beneficial effect or therapeutic benefit to a subject, or anyobjective or subjective transient or temporary, or longer-termimprovement (e.g., cure) in the condition. Thus, a satisfactory clinicalendpoint is achieved when there is an incremental improvement in thesubjects condition or a partial reduction in the severity, frequency,duration or progression of one or more associated adverse symptoms orcomplications or inhibition, reduction, elimination, prevention orreversal of one or more of the physiological, biochemical or cellularmanifestations or characteristics of the condition, disorder or disease.A therapeutic benefit or improvement (“ameliorate” is used synonymously)therefore need not be complete ablation of any or all adverse symptomsor complications associated with the condition, disorder or disease butis any measurable or detectable objectively or subjectively meaningfulimprovement in the condition, disorder or disease. For example,inhibiting a worsening or progression of the condition, disorder ordisease, or an associated symptom (e.g., slowing or stabilizing one ormore symptoms, complications or physiological or psychological effectsor responses), even if only for a few days, weeks or months, even ifcomplete ablation of the condition, disorder or disease, or anassociated adverse symptom is not achieved is considered to bebeneficial effect.

Prophylactic methods are included. “Prophylaxis” and grammaticalvariations thereof mean a method in accordance with the invention inwhich contact, administration or in vivo delivery to a subject is priorto manifestation or onset of a condition, disorder or disease (or anassociated symptom or physiological or psychological response), suchthat it can eliminate, prevent, inhibit, decrease or reduce theprobability, susceptibility, onset or frequency of having a condition,disorder or disease, or an associated symptom. Target subject's forprophylaxis can be one of increased risk (probability or susceptibility)of contracting the condition, disorder or disease, or an associatedsymptom, or recurrence of a previously diagnosed condition, disorder ordisease, or an associated symptom, as set forth herein.

Any compound or agent (e.g., drug), therapy or treatment having abeneficial, additive, synergistic or complementary activity or effect(beneficial or therapeutic) can be used in combination with a bindingagent in accordance with the invention. A “second compound” or “secondagent” refers to any compound or agent (e.g., drug) that is not thefirst compound or agent of the recited composition, e.g., if a firstdrug or agent is a particular LIGHT inhibitor, then a second drug oragent is different from the first LIGHT inhibitor. The second compoundor agent can but need not be selective, for example, for binding toLIGHT, HVEM or LTβR.

In accordance with the invention there are provided methods in which asecond compound or agent (e.g., drug) is administered to the subject. Inone embodiment, a second compound or agent (e.g., drug) is administeredto the subject prior to, with or following contacting or administering aLIGHT inhibitor.

Methods of the invention therefore include combination therapies andtreatments. Examples of such combination therapies include separate orpooled compounds or LIGHT inhibitors (e.g., pooled antagonists,compounds or agents). Accordingly, combination compositions, therapiesand treatments are provided, as well as methods of using suchcombinations, therapies and treatments in conjunction with the methodsof the invention. Contact, administration or in vivo delivery of acompound or agent, such as a binding agent, or practice of a therapy ortreatment, can occur prior to, in conjunction with or following a methodor method step of the invention, e.g., prior to, in conjunction orfollowing administering a LIGHT inhibitor.

Non-limiting examples of functional classes of compounds and agentsuseful as a second compound or agent (e.g., drug) includeanti-inflammatory, anti-asthmatic, airway dilators (e.g., xanthine drugssuch as methylxanthines, which are broncho-dilators) and anti-allergydrugs. Additional non-limiting examples of compounds and agents usefulfor employing in the invention, for example to treat an allergiccondition, disorder or disease (e.g., asthma, allergic rhinitis) includehormones, such as steroids (e.g., glucocorticoids); antihistamines; betaadrenergic agonists; anticholinergics; methylxanthines; anti-IgE;anti-leukotrienes; anti-beta2 integrins; anti-alpha-4 integrins;H1-receptor antagonists; anti-CCR3 antagonists; and anti-selectins.

Specific non-limiting examples of glucocorticoids include dexamethasone,triamcinolone acetonide (AZMACORT®), beclomethasone, dipropionate(VANCERIL®), flunisolide (AEROBID®), fluticasone propionate (FLOVENT®),prednisone, methylprednisolone and mometasonefuroate (ASMANEX®,TWISTHALER®). Specific non-limiting examples of antihistamines includechlorcyclizine, chlorpheniramine, triprolidine (ACTIFED®),diphenhydramine hydrochloride (BENADRYL®), fexofenadine hydrochloride(ALLEGRA®), hydroxyzine hydrochloride (ATARAX®), loratadine (CLARITIN®),promethazine hydrochloride (PHENERGAN®), pyrilamine; and anti-IgEomalizumab (XOLAIR®). Specific non-limiting example of beta adrenergicagonists include albuterol (VENTOLIN®; PROVENTIL®), Xopenex®, (S)-isomersubtracted from racemic albuterol (Sepracor Inc.), pirbuterol,epinephrine, racepinephrine, adrenaline, isoproterenol, salmeterol(Serevent®), metaproterenol (ALUPENT®), bitolterol (Tornalate®),fenoterol (BEROTEC®), formoterol (Foradil®), isoetharine, procaterol,β2-adrenoceptor and terbutaline (BRETHINE®, LAMISIL®). A specificnon-limiting example of an anticholinergic (cholinergic receptorantagonist) includes ipratropium bromide (ATROVENT®) and tiotropium.Specific non-limiting examples of methylxanthines include theophylline,aminophylline, theobromine, cromolyn (Intal®) and nedocromil (Fisons). Aspecific non-limiting example of an anti-IgE is omalizumab (XOLAIR®).Specific non-limiting examples of anti-leukotrienes (leukotrieneinhibitors) include cysteinyl-leukotriene (Cys-LT), Singulair® andAccolate®.

Anti-inflammatory agents useful for employing in the methods includecytokines and chemokines. Particular non-limiting examples of cytokinesinclude anti-inflammatory cytokines such as IL-4 and IL-10.Anti-cytokines and anti-chemokines, such as antibodies that bind topro-inflammatory cytokines, TNFα, IFNγ, IL-1, IL-2, IL-6, etc., as wellas anti-Th2 cytokines such as IL-5, IL-13, etc., can be employed in themethods.

Additional functional classes of compounds and agents useful as a secondcompound or agent (e.g., drug) include selective or non-selectivepotassium channel activators (bronchodilatators); muscarinic M3 receptorantagonists; M2 receptor agonists; opioid receptor agonists (inhibitrelease of sensory neuropeptides); H3-receptor agonists (inhibitacetylcholine release); phospholipase A2 inhibitors; 5-lipoxygenaseinhibitors; 5-lipoxygenase activating protein (FLAP) inhibitors;phosphodiesterase inhibitors; immunomodulating agents (Ciclosporine);antibody against adhesion molecules; and antagonists of tachykinins(e.g., Substance P or neurokinin).

Compositions including LIGHT inhibitors can be included in apharmaceutically acceptable carrier (excipient, diluent, vehicle orfilling agent) for administration to a subject. The terms“pharmaceutically acceptable” and “physiologically acceptable” mean abiologically acceptable formulation, gaseous, liquid or solid, ormixture thereof, which is suitable for one or more routes ofadministration, in vivo delivery or contact. Such formulations includesolvents (aqueous or non-aqueous), solutions (aqueous or non-aqueous),emulsions (e.g., oil-in-water or water-in-oil), suspensions, syrups,elixirs, dispersion and suspension media, coatings, isotonic andabsorption promoting or delaying agents, compatible with pharmaceuticaladministration or in vivo contact or delivery. Aqueous and non-aqueoussolvents, solutions and suspensions may include suspending agents andthickening agents. Such pharmaceutically acceptable carriers includetablets (coated or uncoated), capsules (hard or soft), microbeads,powder, granules and crystals.

Cosolvents and adjuvants may be added to the formulation. Non-limitingexamples of cosolvents contain hydroxyl groups or other polar groups,for example, alcohols, such as isopropyl alcohol; glycols, such aspropylene glycol, polyethyleneglycol, polypropylene glycol, glycolether; glycerol; polyoxyethylene alcohols and polyoxyethylene fatty acidesters. Adjuvants include, for example, surfactants such as, soyalecithin and oleic acid; sorbitan esters such as sorbitan trioleate; andpolyvinylpyrrolidone.

Supplementary active compounds (e.g., preservatives, antioxidants,antimicrobial agents including biocides and biostats such asantibacterial, antiviral and antifungal agents) can also be incorporatedinto the compositions. Pharmaceutical compositions may therefore includepreservatives, anti-oxidants and antimicrobial agents.

Preservatives can be used to inhibit microbial growth or increasestability of the active ingredient thereby prolonging the shelf life ofthe pharmaceutical formulation. Suitable preservatives are known in theart and include, for example, EDTA, EGTA, benzalkonium chloride orbenzoic acid or benzoates, such as sodium benzoate. Antioxidantsinclude, for example, ascorbic acid, vitamin A, vitamin E, tocopherols,and similar vitamins or provitamins.

An antimicrobial agent or compound directly or indirectly inhibits,reduces, delays, halts, eliminates, arrests, suppresses or preventscontamination by or growth, infectivity, replication, proliferation,reproduction, of a pathogenic or non-pathogenic microbial organism.Classes of antimicrobials include, antibacterial, antiviral, antifungaland antiparasitics. Antimicrobials include agents and compounds thatkill or destroy (-cidal) or inhibit (-static) contamination by orgrowth, infectivity, replication, proliferation, reproduction of themicrobial organism.

Exemplary antibacterials (antibiotics) include penicillins (e.g.,penicillin G, ampicillin, methicillin, oxacillin, and amoxicillin),cephalosporins (e.g., cefadroxil, ceforanid, cefotaxime, andceftriaxone), tetracyclines (e.g., doxycycline, chlortetracycline,minocycline, and tetracycline), aminoglycosides (e.g., amikacin,gentamycin, kanamycin, neomycin, streptomycin, netilmicin, paromomycinand tobramycin), macrolides (e.g., azithromycin, clarithromycin, anderythromycin), fluoroquinolones (e.g., ciprofloxacin, lomefloxacin, andnorfloxacin), and other antibiotics including chloramphenicol,clindamycin, cycloserine, isoniazid, rifampin, vancomycin, aztreonam,clavulanic acid, imipenem, polymyxin, bacitracin, amphotericin andnystatin.

Particular non-limiting classes of anti-virals include reversetranscriptase inhibitors; protease inhibitors; thymidine kinaseinhibitors; sugar or glycoprotein synthesis inhibitors; structuralprotein synthesis inhibitors; nucleoside analogues; and viral maturationinhibitors. Specific non-limiting examples of anti-virals includenevirapine, delavirdine, efavirenz, saquinavir, ritonavir, indinavir,nelfinavir, amprenavir, zidovudine (AZT), stavudine (d4T), larnivudine(3TC), didanosine (DDI), zalcitabine (ddC), abacavir, acyclovir,penciclovir, valacyclovir, ganciclovir,1,-D-ribofuranosyl-1,2,4-triazole-3 carboxamide, 9->2-hydroxy-ethoxymethylguanine, adamantanamine, 5-iodo-2′-deoxyuridine,trifluorothymidine, interferon and adenine arabinoside.

Exemplary antifungals include agents such as benzoic acid,undecylenicalkanolamide, ciclopiroxolamine, polyenes, imidazoles,allylamine, thicarbamiates, amphotericin B, butylparaben, clindamycin,econaxole, amrolfine, butenafine, naftifine, terbinafine, ketoconazole,elubiol, econazole, econaxole, itraconazole, isoconazole, miconazole,sulconazole, clotrimazole, enilconazole, oxiconazole, tioconazole,terconazole, butoconazole, thiabendazole, voriconazole, saperconazole,sertaconazole, fenticonazole, posaconazole, bifonazole, fluconazole,flutrimazole, nystatin, pimaricin, amphotericin B, flucytosine,natamycin, tolnaftate, mafenide, dapsone, caspofungin, actofunicone,griseofulvin, potassium iodide, Gentian Violet, ciclopirox,ciclopiroxolamine, haloprogin, ketoconazole, undecylenate, silversulfadiazine, undecylenic acid, undecylenicalkanolamide andCarbol-Fuchsin.

The pH can be adjusted by use or addition of pharmacologicallyacceptable acids or bases. Examples of inorganic acids include:hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and/orphosphoric acid. Examples of organic acids are: ascorbic acid, citricacid, malic acid, tartaric acid, maleic acid, succinic acid, fumaricacid, acetic acid, formic acid and/or propionic acid, etc. Acids whichform an acid addition salt with the active ingredient may also be used.Examples of bases include alkali metal hydroxides and alkali metalcarbonates. If such bases are used, the resulting salts which arecontained in the pharmaceutical formulation, are typically compatiblewith the acid. If desired, mixtures of acids or bases may also be used.

Pharmaceutical compositions can optionally be formulated to becompatible with a particular route of administration. Thus,pharmaceutical compositions include carriers (excipients, diluents,vehicles or filling agents) suitable for administration by variousroutes and delivery to targets, locally, regionally or systemically.

Exemplary routes of administration for contact or in vivo delivery whicha composition can optionally be formulated include respiratory system(nasal, inhalation, respiration, intubation, intrapulmonaryinstillation), oral, buccal, intrapulmonary, intrauterine, intradermal,topical, dermal, parenteral, sublingual, subcutaneous, intravascular,intrathecal, intraarticular, intracavity, transdermal, iontophoretic,intraocular, ophthalmic, optical, intravenous, intramuscular,intraglandular, intraorgan, intralymphatic.

Nasal and instillation formulations typically include aqueous solutionsof active ingredient (compounds or agents) optionally with one or morepreservative or isotonic agents. Such formulations are typicallyadjusted to a pH and isotonic state compatible with nasal mucousmembranes. A solvent may include only water, or it may be a mixture ofwater and one or more other components (e.g., ethanol). Typically, themaximum ethanol is up to about 70-75%% by volume. The remaining volumemay consist of water or one or more other solvents in variousproportions.

Formulations that include respirable or inhalable liquid or solidparticles of the active ingredient (e.g., compound, binding agent) canhave particles of a size sufficiently small to pass through the mouthand larynx upon inhalation and continue into the airways of the lungs(e.g., bronchi and alveoli). Particles typically range in size fromabout 0.05, about 0.1, about 0.5, about 1, about 2 to about 4, about 6,about 8, about 10 microns in diameter. Particles of non-respirable sizecan be included in an aerosol or spray to deposit in the throat. Fornasal administration or intrapulmonary instillation, a particle size inthe range of about 8, about 10, about 20, about 25 to about 35, about50, about 100, about 150, about 250, about 500 μm (diameter) is typicalfor retention in nasal cavity or for instillation into lung.

Formulations suitable for parenteral administration comprise aqueous andnon-aqueous solutions, suspensions or emulsions of the active compound,which preparations are typically sterile and can be isotonic with theblood of the intended recipient. Non-limiting illustrative examplesinclude water, saline, dextrose, fructose, ethanol, animal, vegetable orsynthetic oils.

For transmucosal or transdermal administration (e.g., topical contact),penetrants can be included in the pharmaceutical composition. Penetrantsare known in the art, and include, for example, for transmucosaladministration, detergents, bile salts, and fusidic acid derivatives.For transdermal administration, the active ingredient can be formulatedinto aerosols, sprays, ointments, salves, gels, or creams as generallyknown in the art. For contact with skin, pharmaceutical compositionstypically include ointments, creams, lotions, pastes, gels, sprays,aerosols, or oils. Carriers which may be used include Vaseline, lanolin,polyethylene glycols, alcohols, transdermal enhancers, and combinationsthereof.

Compounds including LIGHT inhibitors, either alone or in combinationwith a pharmaceutically acceptable carrier, second compound, drug, etc.can be administered into the respiratory system of a subject byinhalation, respiration, intubation, or intrapulmonary instillation(into the lungs), for example. Respiratory administration can beachieved using an aerosol or spray of a gas, liquid or powdered nasal,intrapulmonary, respirable or inhalable in a particle form. Theparticles include the compound or binding agent, and optionally anyother component (e.g., second compound), and are administered ordelivered to the subject by inhalation, by nasal administration orinstillation into the airways or the lung.

Administration to airways can be accomplished using an article ofmanufacture, such as container with or without an aerosol. Liquidformulations may be squirted into the respiratory system (e.g., nose)and the lung from a container by pressure or using an aerosol propellantor a spray device or delivery system. Administration can be passive orit can be assisted by a pressurized delivery system or device. Anaerosol, delivery system or device can include a pressurized containercontaining liquid, gas or dry powder.

An “aerosol formulation” refers to a preparation that includes dropletsor particles of active ingredient (e.g., compound, binding agent)suitable for delivery to respiratory system (e.g., lung, airway, nasaland sinus epithelium). The aerosol formulation can include a sufficientor effective amount of a compound or agent and a pharmaceuticallyacceptable carrier, optionally a propellant, in a container or aerosolor spray device or delivery system. Aerosol formulations can deliverhigh concentrations into airways with relatively low systemicabsorption, and include for example nasal sprays, inhalation solutions,inhalation suspensions, and inhalation sprays. Nasal sprays typicallycontain active ingredient dissolved or suspended in solution or in anexcipient, in nonpressurized dispensers that deliver a metered dose ofthe ingredient.

For aerosol delivery, pH of the formulation is typically between 5.0 and7.0. If the aerosol is too acidic or basic, it can cause bronchospasmand cough. The tolerized pH range is relative and depends on a patient'stolerance: some patients tolerate a mildly acidic aerosol, which inothers will cause bronchospasm. Typically, an aerosol formulation havinga pH less than 4.5 induces bronchospasm.

Compositions including compounds and binding agents can be formulated ina dry powder for delivery into the endobronchial space. Dry powderformulations provide stability, high volume delivery per puff, and lowsusceptibility to microbial growth. Dry powder formulations typicallyare stable at ambient temperature, and have a physiologically acceptablepH of 4.0-7.5. Dry powder formulations are convenient because they donot require any further handling, such as dilution, prior toadministration. Depending on delivery device efficiency, effective drypowder dosage levels typically fall in the range of about 10 to about100 mg. Dry powder formulations can be used directly in metered dose ordry powder inhalers.

Aerosol and spray delivery systems and devices, also referred to as“aerosol generators” and “spray generators” are known in the art andinclude metered dose inhalers (MDI), nebulizers (ultrasonic, electronicand other nebulizers), nasal sprayers and dry powder inhalers.

MDIs typically include an actuator, a metering valve, and a containerthat holds a suspension or solution, propellant, and surfactant (e.g.,oleic acid, sorbitan trioleate, lecithin). The container may bepressurized or not, but typically it is either squeezed to dispense theingredient, or has an actuator connected to a metering valve so thatactivation of the actuator causes a predetermined amount to be dispensedfrom the container in the form of an aerosol, which is inhaled by thesubject. MDIs typically use liquid propellant. Typically, metered-doseaerosol inhalers create droplets that are 15 to 30 microns in diameter.Currently, MDI technology is optimized to deliver masses of 1 microgramto 10 mg of a therapeutic.

Nebulizers, also referred to as atomizers, are devices that turnmedication into a fine mist inhalable by a subject through a face maskthat covers the mouth and nose. Nebulizers provide small droplets andhigh mass output which can be delivered to upper and lower respiratoryairways. Typically, nebulizers create droplets down to about 1 micron indiameter. Doses administered by nebulizers are typically larger thandoses administered by MDIs.

Nebulizers include air-jet and ultrasonic nebulizers, in fluidconnection with a reservoir containing disposed therein a solution orsuspension of active ingredient. Nebulizers (air-jet, ultrasonic orelectronic) are typically used for acute care of nonambulatory patientsand in infants and children. Airjet nebulizers are relatively large butconsidered portable because of the availability of small compressed airpumps. Ultrasonic and electronic nebulizers are typically more portablebecause they usually do not require a source of compressed air. Anexample of an airjet nebulizer is the NE-C25 CompAir XLT CompressorNebulizer System (Omron® Healthcare). Examples of ultrasonic nebulizersinclude the Zewa Portable Ultrasonic Nebulizer (Zewa, Inc.); the MabisMist II Ultrasonic Nebulizer (Mabis Healthcare, Inc.); and the MICROAirUltrasonic Nebulizer (Omron® Healthcare). An example of an electronicnebulizer is the Micro-Air® Electronic Nebulizer with V.M.T. (Omron®Healthcare). Modified nebulizers can have the addition of a one-way flowvalve (e.g., Pari LC Plus™, Pari Respiratory Equipment, Inc.), whichdelivers up to 20% more drug than unmodified nebulizers.

Components of the nebulizer are typically made of a material suitablefor their intended function. The housing of the nebulizer and, if thefunction allows, other parts can be made of plastic (PVC, Polycarbonate,polystyrene, polypropylene, polybutylene, etc.). Plastic can be formedby injection molding. For medical applications, physiologicallyacceptable materials are used.

Dry-powder inhalers (DPI) can be used to deliver the compounds oragents, either alone or in combination with a pharmaceuticallyacceptable carrier, second compound, etc. Dry powder inhalers deliveractive ingredient to airways and lungs while the subject inhales throughthe device. DPIs typically do not contain propellants or any otheringredients, only the medication, but may optionally include othercomponents. DPIs are typically breath-activated, but may involve air orgas pressure to assist delivery. For breath-activated DPIs, a subjectneed not coordinate breathing with the activation of the inhaler.

There are two major DPI design classes. The first is a device-meteringdesign in which a reservoir of drug is stored within the device and thesubject ‘loads’ a dose of the device into the inhalation chamber, andthe inspiratory flow of the patient accelerates the powder out of thedevice and into the oral cavity. The second type of DPI may also employan air source, a gas source, or electrostatics, in order to deliver theactive ingredient. Non-limiting examples of DPIs include Spinhaler®(Rhone-Poulenc Rorer Pharmaceuticals, Collegeville, Pa.), Inhalator®(Boehringer Ingelheim, Ingelheim, Germany), Rotahaler®(GlaxoSmithKline), Turbulaler® (Astra Draco Pharmaceuticals, Lund,Sweden) and Accuhaler (GlaxoSmithKline).

An aerosol, delivery system or device can include a propellant.Exemplary propellants include chlorofluorocarbons (e.g.,trichlorofluoromethane, dichlorodifluoromethane,dichlorotetrafluoromethane, CFC-11, CFC-12) and the non-dchlorofluorocarbons, HFC-134A and HFC-227. Suitable fluorocarbon (HFA)propellants are known in the art and include, for example, HFA134a(1,1,1,2-tetrafluoroethane), HFA227(1,1,1,2,3,3,3-heptafluoro-n-propane) and mixtures of HFA134a andHFA227.

Pharmaceutical compositions and delivery systems appropriate forcompositions and methods of the invention are known to the skilledartisan (see, e.g., Remington: The Science and Practice of Pharmacy(2003) 20^(th) ed., Mack Publishing Co., Easton, Pa.; Remington'sPharmaceutical Sciences (1990) 18^(th) ed., Mack Publishing Co., Easton,Pa.; The Merck Index (1996) 12^(th) ed., Merck Publishing Group,Whitehouse, N.J.; Pharmaceutical Principles of Solid Dosage Forms(1993), Technonic Publishing Co., Inc., Lancaster, Pa.; Ansel andStoklosa, Pharmaceutical Calculations (2001) 11^(th) ed., LippincottWilliams & Wilkins, Baltimore, Md.; and Poznansky et al., Drug DeliverySystems (1980), R. L. Juliano, ed., Oxford, N.Y., pp. 253-315)

LIGHT inhibitors and pharmaceutical compositions thereof can be packagedin unit dosage form (capsules, troches, cachets, lozenges, or tablets)for ease of administration and uniformity of dosage. “Unit dosage form”as used herein refers to physically discrete units suited as dosages fortreatment or therapy. Each unit contains a predetermined quantity ofagent in association with the pharmaceutical carrier (excipient,diluent, vehicle or filling agent) which, when administered in one ormore doses, is calculated to produce a desired beneficial effect. Unitdosage forms also include, for example, ampules and vials, which mayinclude a composition in a freeze-dried or lyophilized state; a sterileliquid carrier, for example, can be added prior to administration ordelivery in vivo. Unit dosage forms additionally include, for example,ampules and vials with liquid compositions disposed therein. Unit dosageforms further include compositions for transdermal administration, suchas “patches” adapted to remain in contact with the epidermis of theintended recipient for an extended or brief period of time. Theindividual unit dosage forms can be included in multi-dose kits orcontainers.

Dose amounts, frequency and duration for binding agents, including LIGHTinhibitors, or pro-drugs thereof, can be can be empirically determinedin appropriate animal models. Dose amounts, frequency and duration canalso be determined and optimized in human clinical trials.

The dosage amount can range from about 0.0001 mg/kg of subject bodyweight/day to about 1,000.0 mg/kg of subject body weight/day. Of course,doses can be more or less, as appropriate, for example, 0.00001 mg/kg ofsubject body weight to about 10,000.0 mg/kg of subject body weight,about 0.001 mg/kg, to about 1,000 mg/kg, about 0.01 mg/kg, to about 100mg/kg, or about 0.1 mg/kg, to about 10 mg/kg of subject body weight overa given time period, e.g., 1, 2, 3, 4, 5 or more hours, days, weeks,months, years, in single bolus or in divided/metered doses.

As a non-limiting example, for treatment of lung or airway inflammation,or asthma (e.g., allergic or non-allergic asthma or rhinitis), a subjectmay be administered in single bolus or in divided/metered doses in therange of about 10 to 50,000 micrograms (“mcg”)/day, 10 to 20,000mcg/day, 10 to 10,000 mcg/day, 25-1,000 mcg/day, 25 to 400 mcg/day,25-200 mcg/day, 25-100 mcg/day or 25-50 mcg/day, which can be adjustedto be greater or less according to the weight of the subject, e.g., perpound, kilogram, etc.

LIGHT inhibitors, combinations of LIGHT inhibitors and other actives andpharmaceutical formulations thereof can be administered to a subject atany frequency, as a single bolus or in divided/metered doses, one, two,three, four or more times over a given time period, e.g., per hour, day,week, month or year. Exemplary dosage frequencies for airway or lungconditions, disorders or diseases, such as asthma can vary, but aretypically from 1-7 times, 1-5 times, 1-3 times, 2-times or once, daily,weekly or monthly, to reduce, inhibit, decrease, delay, prevent, halt oreliminate progression, severity, frequency, duration, or probability ofone or more adverse symptoms of the conditions, disorders or diseases,as set forth herein or that would be apparent to one skilled in the art.Timing of contact, administration or in vivo delivery can be dictated bythe condition, disorder or disease to be treated. For example, an amountcan be administered to the subject substantially contemporaneously with,or within about 1-60 minutes or hours of the onset of a symptomassociated with or caused by lung or airway inflammation, asthma (e.g.,non-allergic asthma, allergic asthma, or an asthmatic episode) orairway-constriction, -narrowing or -obstruction, or a respiratory,interstitial or pulmonary disease or disorder.

Dosage amount, frequency or duration can be increased, if necessary, orreduced, for example, once control of the condition, disorder or diseaseis achieved, dose amounts, frequency or duration can be reduced. Otherconditions, disorders or diseases of the airways and lungs can besimilarly treated, dosing amount, frequency or duration reduced, whenadequate control of the condition, disorder or disease is achieved.

Of course, the dosage amount, frequency and duration can vary dependingupon the judgment of the skilled artisan which will consider variousfactors such as whether the treatment is prophylactic or therapeutic,the type or severity of the condition, disorder or disease, theassociated symptom to be treated, the clinical endpoint(s) desired suchas the type and duration of beneficial or therapeutic effect. Additionalnon-limiting factors to consider in determining appropriate dosageamounts, frequency, and duration include previous or simultaneoustreatments, potential adverse systemic, regional or local side effects,the individual subject (e.g., general health, age, gender, race,bioavailability), condition of the subject such as other disorders ordiseases present and other treatments or therapies that the subject hasor is undergoing (e.g., medical history). The skilled artisan willappreciate the factors that may influence the dosage, frequency andduration required to provide an amount sufficient to provide a subjectwith a beneficial effect, such as a therapeutic benefit.

The invention provides kits including LIGHT inhibitors suitable forpracticing the methods, treatment protocols or therapeutic regimesherein, and suitable packing material. In one embodiment, a kit includesa LIGHT inhibitor, and instructions for administering or using the LIGHTinhibitor. In another embodiment, a kit includes a LIGHT inhibitor, anarticle of manufacture for delivery of the LIGHT inhibitor to the targetarea, organ, tissue or system (e.g., lungs or airways), and instructionsfor administering the LIGHT inhibitor.

The term “packing material” refers to a physical structure housing acomponent of the kit. The material can maintain the componentssterilely, and can be made of material commonly used for such purposes(e.g., paper, corrugated fiber, glass, plastic, foil, ampules, vials,tubes, etc.).

Kits of the invention can include labels or inserts. Labels or insertsinclude “printed matter,” e.g., paper or cardboard, or separate oraffixed to a component, a kit or packing material (e.g., a box), orattached to a ampule, tube or vial containing a kit component. Labels orinserts can additionally include a computer readable medium, such as adisk (e.g., floppy diskette, ZIP disk), optical disk such as CD- orDVD-ROM/RAM, DVD, MP3, magnetic tape, or an electrical storage mediasuch as RAM and ROM or hybrids of these such as magnetic/optical storagemedia, FLASH media or memory type cards.

Labels or inserts can include identifying information of one or morecomponents therein (e.g., the binding agent or pharmaceuticalcomposition), dose amounts, clinical pharmacology of the active agent(s)including mechanism of action, pharmacokinetics and pharmacodynamics.Labels or inserts can include information identifying manufacturerinformation, lot numbers, and location and date of manufacture.

Labels or inserts can include information on a condition, disorder ordisease for which a kit component may be used. Labels or inserts caninclude instructions for the clinician or subject for using one or moreof the kit components in a method, or treatment protocol or therapeuticregimen. Instructions can include dosage amounts, frequency or duration,and instructions for practicing any of the methods, treatment protocolsor therapeutic regimes described herein.

Labels or inserts can include information on any benefit that acomponent may provide, such as a therapeutic benefit. For example, anon-limiting examples of a benefit would be improved breathing orrespiration, increased or improved airway dilation, etc. A benefit couldalso include a reduced need (amount, frequency or duration) for othermedications, treatment protocols or therapeutic regimes that the subjectmay be using or have used for treatment of the condition, disorder ordisease.

Labels or inserts can include information on potential adverse sideeffects, such as warnings to the subject or clinician regardingsituations where it would not be appropriate to use a particularcomposition (e.g., a LIGHT inhibitor). For example, adverse side effectsare generally more likely to occur at higher dose amounts, frequency orduration of the active agent and, therefore, instructions could includerecommendations against higher dose amounts, frequency or duration.Adverse side effects could also occur when the subject has, will be oris currently taking one or more other medications that may beincompatible with the composition, or the subject has, will be or iscurrently undergoing another treatment protocol or therapeutic regimenwhich would be incompatible with the composition and, therefore,instructions could include information regarding such incompatibilities.Non-limiting examples of adverse side effects include, for example,hypersensitivity, rash, neurological effects such as tachycardia;palpitations; headache; tremor and nervousness.

In accordance with the invention, there are provided methods ofidentifying an agent that reduces or inhibits lung or airwayinflammation, methods of identifying an agent for treating asthma andmethods of identifying an agent for treating fibrosis. In oneembodiment, a method includes administering a test inhibitor of LIGHT(p30 polypeptide) to a subject; and measuring lung or airwayinflammation in the subject, wherein a reduction or inhibition of lungor airway inflammation identifies the test inhibitor as an agent thatreduces or inhibits lung or airway inflammation. In another embodiment,a method includes administering a test inhibitor of LIGHT (p30polypeptide) to a subject; and measuring a symptom of asthma in thesubject, wherein a reduction or inhibition of a symptom of asthmaidentifies the test inhibitor as an agent for treating asthma. In afurther embodiment, a method includes administering a test inhibitor ofLIGHT (p30 polypeptide) to a subject; and measuring a symptom offibrosis in the subject, wherein a reduction or inhibition of a symptomof fibrosis identifies the test inhibitor as an agent for treatingfibrosis.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention relates. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described herein.

All applications, publications, patents and other references, GenBankcitations and ATCC citations cited herein are incorporated by referencein their entirety. In case of conflict, the specification, includingdefinitions, will control.

As used herein, the singular forms “a”, “and,” and “the” include pluralreferents unless the context clearly indicates otherwise. Thus, forexample, reference to “a LIGHT inhibitor” includes a plurality of LIGHTinhibitors; and reference to “a symptom” includes a plurality ofsymptoms (e.g., adverse or undesirable). Of course, this does notpreclude limiting certain embodiments of the invention to specific LIGHTinhibitors or antagonists, particular symptoms, particular conditions,disorders or diseases, particular subjects, etc., using appropriatelanguage.

As used herein, all numerical values or numerical ranges includeintegers within such ranges and fractions of the values or the integerswithin ranges unless the context clearly indicates otherwise. Thus, toillustrate, reference to a range of 90-100%, includes 91%, 92%, 93%,94%, 95%, 95%, 97%, etc., as well as 91.1%, 91.2%, 91.3%, 91.4%, 91.5%,etc., 92.1%, 92.2%, 92.3%, 92.4%, 92.5%, etc., and so forth. Referenceto a range of 0-72 hrs, includes 1, 2, 3, 4, 5, 6, 7 hrs, etc., as wellas 1, 2, 3, 4, 5, 6, 7 minutes, etc., and so forth. Reference to a rangeof doses, such as 0.11 ug/kg, 1-10 ug/kg, 10-25 ug/kg, 25-50 ug/kg,50-100 ug/kg, 100-500 ug/kg, 500-1,000 ug/kg, 1-5 mg/kg, 5-10 mg/kg,10-20 mg/kg, 20-50 mg/kg, 50-100 mg/kg, 100-250 mg/kg, 250-500 mg/kg,includes 0.11-0.9 ug/kg, 2-9 ug/kg, 11.5-24.5 ug/kg, 26-49 ug/kg, 55-90ug/kg, 125-400 ug/kg, 750-800 ug/kg, 1.1-4.9 mg/kg, 6-9 mg/kg, 11.5-19.5mg/kg, 21-49 mg/kg, 55-90 mg/kg, 125-200 mg/kg, 275.5-450.1 mg/kg, etc.

The invention is generally disclosed herein using affirmative languageto describe the numerous embodiments. The invention also specificallyincludes embodiments in which particular subject matter is excluded, infull or in part, such as substances or materials, method steps andconditions, protocols, procedures, assays or analysis disclosed herein.As an example, the invention includes embodiments in which specificsubject matter disclosed herein is excluded from the embodiments. Thus,even though the invention is generally not expressed herein in terms ofwhat the invention does not include, aspects that are not expresslyincluded in the invention are nevertheless expressly or inherentlydisclosed herein.

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention.Accordingly, the following examples are intended to illustrate but notlimit the scope of invention described in the claims.

EXAMPLES Example 1

This example includes studies showing LIGHT plays a role in thedevelopment of chronic allergic lung inflammation and airway remodeling.

Wild type C57BL/6 and LIGHT-deficient (LIGHT−/−) mice were sensitizedwith 50 ug ovalbumin (OVA) with 0.5 mg alum intraperitoneally (i.p.) ondays 0 and 12 followed by 20 ug OVA intranasal challenges on days 24,26, 28 and then two times per week for 4 weeks. Percent of BALeosinophils at 1 day and 3 days after last OVA challenge was determined.Results are shown in FIG. 1. BAL eosinophils are from 4 mice per groupfor each time point.

Another study was conducted using wild type C57BL/6 and LIGHT-deficient(LIGHT −/−) mice sensitized with 50 ug ovalbumin (OVA) with 0.5 mg alumintraperitoneally (i.p.) on days 0 and 12 followed by 20 ug intranasalOVA challenges on days 24, 26, 28 and then two times per week for 6weeks. Lung sections were stained with trichrome to measure fibrosis,and for alpha-smooth muscle actin to measure development of smoothmuscle mass, both features of airway remodeling; histographs are shownin FIG. 2A. The area of fibrosis or smooth muscle was quantified usingimage analysis with normalization for bronchial size, as shown in FIG.2B. Results are means +/−SEM of 36-48 bronchial regions per group.

Additionally naïve CD4 cells from wild-type (wt) or LIGHT−/− OT-II TCRtransgenic mice were adoptively transferred into wt B6.PL recipients.Mice were immunized i.p. with OVA in Alum to generate Th2 cells.Transferred OT-II cells were visualized on day 8 by staining for Thy1.2and CD4, as shown in FIG. 3A and FIG. 3C. The degree of apoptosisoccurring within these populations was determined by co-staining forannexin V and 7-AAD after gating on Thy1.2+ cells, as shown in FIG. 3Band FIG. 3D. The percentages of early and late apoptotic cells areindicated. Data are representative of 4 mice per group.

As described above, when LIGHT-deficient mice were sensitized withovalbumin (OVA) followed by repetitive intranasal OVA challenges forfour to six weeks, they exhibited reduced eosinophilic lung inflammation(FIG. 1). Additionally, markedly reduced peribronchial fibrosis andsmooth muscle mass was found in LIGHT-deficient mice. There was a 40-50%reduction in subepithelial fibrosis and smooth muscle mass, importantpathologic remodeling features of chronic asthma (FIG. 2).

LIGHT may be directly acting in the lung to induce chronic asthmaticchanges. A potential source of LIGHT in the inflamed lung are CD4+ Tcells which are recovered at high levels in BAL specimens from humanasthmatics.

To investigate the role of LIGHT on CD4 cells, LIGHT-deficient CD4 Tcells were evaluated in adoptive recipients that were immunized withOVA. When OVA-reactive T cells were enumerated at the peak of theprimary response (day 7), there was a strong reduction in their number(80-90%) when LIGHT was not expressed. As evidence that LIGHT controlsthe longevity and survival of T cells, staining for annexin V and 7-AADin FIGS. 3B and 3D demonstrated that OVA-reactive CD4 cells displayedenhanced percentages undergoing apoptosis, a prelude to death, in theabsence of LIGHT.

Example 2

This example includes studies demonstrating the requirement of LIGHT inlung inflammation by controlling T cells, in a system whereby T cellsthat could not express LIGHT were used to try to induce asthmatic lunginflammation.

Naïve wild-type (wt) or LIGHT−/− OT-II CD4 T cells were activated invitro for 3 days with anti-CD3 and anti-CD28 plus IL-4, IL-2, andanti-IFN-γ, and then recultured without further stimulation for 3 days.The resultant Th2 cells were transferred into naïve B6.PL recipients.Mice were exposed to aerosolized OVA or PBS and then rested for severalweeks. Mice were then re-exposed to aerosolized OVA or PBS for 2consecutive days. 1 day after the last challenge, lung and bronchiallavage samples were obtained. Lung histology, by H&E; sections from twoindividual OVA-challenged mice are shown in FIG. 4A. Total leukocyte andeosinophil counts by differential cytospin stain; and IL-5 and IL-13expression by ELISA, were measured, results are shown in FIG. 4B.

First set of data in each case, animals challenged with PBS. Second set,animals challenged with OVA. Data from individual mice are shown with 3mice per group.

In another study, using the experimental protocol in FIG. 4, the totalnumber of wt OT-II (left two bars) and LIGHT−/− OT-II (right two bars)CD4 T cells accumulating after PBS or OVA challenge were enumerated inmediastinal lymph nodes and lung after staining for CD4 and Thy1.2 (FIG.5). Data were average numbers from 3 mice per group.

In another study, naïve wild-type (Wt) or LIGHT−/− OT-II CD4 T cellswere activated in vitro as in FIG. 4. The Th2 cells were adoptivelytransferred i.v. into naïve C57/BL6 WT hosts. Mice received 20 ug OVAintranasal challenges on day 1, 3, 5 and then chronic airway challengestwo times per week for four weeks beginning one day after cell transferand were sacrificed one day after last challenge. Percentages (left) andabsolute numbers (right) of donor CD4 Th2 cells expressing OVA-specificTCR Vα2Vβ5 were determined in Bronchoalveolar lavage (BAL; FIG. 6A),Lung (FIG. 6B), and lung-draining lymph node (FIG. 6C). Samples werepooled from 3 mice in each group.

Transfer of primed Th2 cells into naïve wild-type recipients followed byadministration of inhaled antigen showed that lung inflammation wasprofoundly impaired when LIGHT was absent (FIG. 4). Inflammatoryinfiltrates in the lung were reduced based on histological examination(FIG. 4A), and the levels of Th2 cytokines and eosinophils present inbronchial lavages were reduced when LIGHT-deficient CD4 cells were used(FIG. 4B). The accumulation of LIGHT-deficient T cells in lungs and lungdraining lymph nodes was substantially lower after airway challenge suchthat a 70-80% reduction in numbers was observed compared to animalsreceiving control T cells that expressed LIGHT (FIG. 5).

To evaluate how LIGHT controls Th2 cells after chronic repetitiveallergen challenges that may be more relevant to human asthma, LIGHTdeficient and wild type CD4 cells were adoptively transferred into wtmice followed by 11 intranasal challenges over 5 weeks. Strikingly,LIGHT-deficient T cells completely failed to accumulate in the lung,bronchoalveolar lavage (BAL), and lung draining lymph nodes (FIG. 6).The data show that LIGHT is essential for controlling T cells in hoststhat are repetitively challenged with antigen in the lung over a longperiod of time.

Example 3

This example includes shows reduction of lung inflammation aftertreatment with lymphotoxin beta receptor fusion protein during chronicallergen challenge via the airways.

Wild type C57BL/6 mice were sensitized with 50 ug ovalbumin (OVA) with0.5 mg alum intraperitoneally (i.p.) on days 0 and 12 followed by 20 ugintranasal OVA challenges on days 24, 26, 28. Mice were thenintranasally challenged two times per week for four more weeks with OVA,and lymphotoxin beta receptor Ig fusion protein (LTβR-Ig) wasadministered i.p. (50 ug) twice per week starting one day before theseOVA challenges. FIG. 7A shows total cell infiltrate and eosinophils inBAL and FIG. 7B shows Total cell infiltrate and eosinophils in lungs.BAL results from 6 mice per group +/−SEM and Lung cell results are frompooled lungs from 3 mice per group.

In order to test a therapeutic intervention in the model of chronicasthma, a fusion protein (as described above) which contains thelymphotoxin beta receptor (LTβR) attached to the constant region ofhuman immunoglobulin IgG (LTβR-Ig) was administered to mice only afteracute OVA-induced lung inflammation was established, and treatmentcontinued for four weeks during repetitive antigen challenges via theairways. Compared with mice that received control IgG there were reducedBAL leukocytes and eosinophils, along with markedly decreased total lungcells and eosinophils (FIG. 7). These results show that treatmentstargeting LIGHT can be efficacious in chronic asthma.

The data show that the TNFR superfamily member LIGHT is critical to thedevelopment of allergen induced airway remodeling and fibrosis.Disruption of the LIGHT pathways by targeting LIGHT therapeuticallyreduces lung inflammation. The foregoing data therefore indicate thattreatment targeting LIGHT can be efficacious in asthma as well as otherfibroproliferative diseases in the lung, and other organs.

1. A method for reducing or inhibiting lung or airway inflammation,comprising administering a sufficient amount of an inhibitor of LIGHT(p30 polypeptide) to a subject in need thereof to reduce or inhibit lungor airway inflammation in the subject.
 2. (canceled)
 3. A method fortreating asthma, comprising administering a sufficient amount of aninhibitor of LIGHT (p30 polypeptide) to a subject in need thereof totreat asthma.
 4. A method for treating a respiratory disease ordisorder, comprising administering a sufficient amount of an inhibitorof LIGHT (p30 polypeptide) to a subject in need thereof to treat therespiratory disease or disorder.
 5. A method for treating aninterstitial or pulmonary disease or disorder, comprising administeringa sufficient amount of an inhibitor of LIGHT (p30 polypeptide) to asubject to treat the interstitial or pulmonary disease or disorder.
 6. Amethod for treating a fibrotic disease or disorder, comprisingadministering a sufficient amount of an inhibitor of LIGHT (p30polypeptide) to a subject to treat the fibrotic disease or disorder. 7.The method of claim 4, wherein the respiratory disease or disorderaffects the upper or lower respiratory tract.
 8. The method of claim 4,wherein the respiratory disease or disorder comprises asthma, allergicasthma, bronchiolitis or pleuritis.
 9. The method of claim 4, whereinthe respiratory disease comprises an allergic disorder.
 10. The methodof claim 9, wherein the allergic disorder is selected from: Extrinsicbronchial asthma; Allergic rhinitis; Onchocercal dermatitis; Atopicdermatitis, Drug reactions; Nodules, eosinophilia, rheumatism,dermatitis, and swelling (NERDS); Eosophageal and a gastrointestinalallergy.
 11. The method of claim 4, wherein the respiratory disease ordisorder is selected from: Airway Obstruction, Apnea, Asbestosis,Atelectasis, Berylliosis, Bronchiectasis, Bronchiolitis, BronchiolitisObliterans Organizing Pneumonia, Bronchitis, Bronchopulmonary Dysplasia,Common Cold, Cough, Empyema, Pleural Empyema, Pleural Epiglottitis,Hemoptysis, Hypertension, Kartagener Syndrome, Meconium Aspiration,Pleural Effusion, Pleurisy, Pneumonia, Pneumothorax, RespiratoryDistress Syndrome, Respiratory Hypersensitivity, Respiratory TractInfections, Rhinoscleroma, Scimitar Syndrome, Severe Acute RespiratorySyndrome, Silicosis, Tracheal Stenosis, Whooping Cough and Influenza.12. The method of claim 5, wherein the interstitial or pulmonary diseaseor disorder, or the fibrotic disease or disorder, is selected from:Eosinophilic pleural effusions; Transient pulmonary eosinophilicinfiltrates (Löffler); Histiocytosis; Chronic eosinophilic pneumonia;Hypersensitivity pneumonitis; Allergic bronchopulmonary aspergillosis;Sarcoidosis; Idiopathic pulmonary fibrosis; pulmonary edema; pulmonaryembolism; pulmonary emphysema; Pulmonary Hyperventilation; PulmonaryAlveolar Proteinosis; Chronic Obstructive Pulmonary Disease (COPD);Interstitial Lung Disease; and Topical eosinophilia.
 13. The method ofany of claims 1 to 12, wherein treatment reduces, decreases, inhibits,delays, eliminates or prevents the probability, severity, frequency, orduration of one or more symptoms associated with or caused by the lungor airway inflammation, asthma or respiratory, interstitial, orpulmonary disease or disorder, or fibrotic disease or disorder.
 14. Themethod of any of claims 1 to 12, wherein one or more symptoms of thelung or airway inflammation, asthma or respiratory, interstitial, orpulmonary disease or disorder, or the fibrotic disease or disorder, isreduced, inhibited, abrogated, eliminated or reversed.
 15. The method ofclaim 14, wherein the symptom comprises shortness of breath (dyspnea),wheezing, stridor, coughing, airway remodeling, rapid breathing(tachypnea), prolonged expiration, runny nose, rapid or increased heartrate (tachycardia), rhonchous lung, over-inflation of the chest orchest-tightness, decreased lung capacity, an acute asthmatic episode, orlung, airway or respiratory mucosum inflammation or tissue damage. 16.The method of claim 14, wherein the symptom comprises infiltration ofeosinophils in lung, lung draining lymph nodes or airway, leukocyteinfiltration of lung draining lymph nodes or airway, hyperplasia ofmucus secreting epithelium, inflammatory lesion of lung, goblet cellhyperplasia, or increased Th2 cytokine production.
 17. The method ofclaim 16, wherein the Th2 cytokine is an interleukin (IL).
 18. Themethod of claim 17, wherein the interleukin (IL) comprises IL-4, IL-5,IL-9, IL-13, IL-16, IL-17 or IL-25.
 19. The method of any of claims 1 to12, wherein the method reduces or inhibits progression, severity,frequency, duration or probability of a symptom of the lung or airwayinflammation, asthma or respiratory, interstitial, or pulmonary diseaseor disorder, or fibrotic disease or disorder.
 20. The method of claims 1to 6, wherein the lung or airway inflammation, asthma or respiratory,interstitial, or pulmonary disease or disorder, or fibrotic disease ordisorder, is caused by an allergen or by exercise.
 21. The method ofclaims 1 to 6, wherein the lung or airway inflammation, asthma orrespiratory, interstitial, or pulmonary disease or disorder, or fibroticdisease or disorder, is chronic or acute.
 22. A method for reducing orinhibiting a Th2-inflammatory response, comprising administering asufficient amount of an inhibitor of LIGHT (p30 polypeptide) to asubject to reduce or inhibit Th2-inflammatory response.
 23. The methodof claim 22, wherein the Th2-inflammatory response affects an organ ortissue of a subject.
 24. A method of reducing or decreasing theprobability, severity, frequency, duration or preventing a subject fromhaving an acute asthmatic episode, comprising administering to a subjectthat has previously experienced an asthmatic episode or has beendiagnosed as having asthma an amount of an inhibitor of LIGHT (p30polypeptide) to reduce or decrease the probability, severity, frequency,duration or prevent an acute asthmatic episode.
 25. The method of claim24, wherein the acute asthmatic episode is caused by or associated withallergic asthma. 26.-27. (canceled)
 28. The method of any of claims 1 to6, 22, 24, 26 and 27, wherein the inhibitor of LIGHT (p30 polypeptide)comprises an antibody or subsequence thereof that binds to LIGHT (p30polypeptide), an antibody or subsequence thereof that binds to HVEM(herpesvirus entry mediator), or an antibody or subsequence thereof thatbinds to LTβR (lymphotoxin beta receptor).
 29. The method of claim 28,wherein the antibody is human or humanized.
 30. The method of any ofclaims 1 to 6, 22, 24, 26 and 27, wherein the inhibitor of LIGHT (p30polypeptide) comprises an LTβR (lymphotoxin beta receptor), HVEM orLIGHT polypeptide subsequence, variant sequence, chimeric sequence ordominant negative sequence.
 31. The method of claim 30, wherein thechimeric sequence comprises a fusion of an LTβR or HVEM polypeptidesequence and an immunoglobulin sequence.
 32. The method of any of claims1 to 6, 22, 24, 26 and 27, wherein the subject is a mammal.
 33. Themethod of any of claims 1 to 6, 22, 24, 26 and 27, wherein the subjectis a human.
 34. The method of any of claims 1 to 6, wherein the methodreduces or decreases undesirable or abnormal eosinophil migration,chemotaxis or generation. 35.-38. (canceled)
 39. The method of any ofclaims 1 to 6, 22, 24, 26 and 27, wherein the inhibitor of LIGHT (p30polypeptide) is administered via inhalation.
 40. The method of any ofclaims 1 to 6, 22, 24, 26 and 27, wherein the inhibitor of LIGHT (p30polypeptide) is formulated into an aerosol. 41.-44. (canceled)
 45. Themethod of any of claims 1 to 6, 22, 24, 26 and 27, wherein the inhibitorof LIGHT (p30 polypeptide) is delivered to the lung or airways.
 46. Amethod of identifying an agent that reduces or inhibits lung or airwayinflammation, comprising: a) administering a test inhibitor of LIGHT(p30 polypeptide) to a subject; and b) measuring lung or airwayinflammation in the subject, wherein a reduction or inhibition of lungor airway inflammation identifies the test inhibitor as an agent thatreduces or inhibits lung or airway inflammation.
 47. A method ofidentifying an agent for treating asthma, comprising: a) administering atest inhibitor of LIGHT (p30 polypeptide) to a subject; and b) measuringa symptom of asthma in the subject, wherein a reduction or inhibition ofa symptom of asthma identifies the test inhibitor as an agent fortreating asthma.
 48. A method of identifying an agent for treatingfibrosis, comprising: a) administering a test inhibitor of LIGHT (p30polypeptide) to a subject; and b) measuring a symptom of fibrosis in thesubject, wherein a reduction or inhibition of a symptom of fibrosisidentifies the test inhibitor as an agent for treating fibrosis.